
==== Front
Hum Vaccin Immunother
Hum Vaccin Immunother
Human Vaccines & Immunotherapeutics
2164-5515
2164-554X
Taylor & Francis

39246041
10.1080/21645515.2024.2394265
2394265
Version of Record
Research Article
Coronavirus
Influence of previous COVID-19 exposure and vaccine type (CoronaVac, ChAdOx1 nCov-19 or BNT162b2) on antibody and cytokine (Th1 or Th2) responses
D. L. PADILLA-BÓRQUEZ ET AL.
HUMAN VACCINES & IMMUNOTHERAPEUTICS
Padilla-Bórquez Diana Lourdes a
Matuz-Flores Mónica Guadalupe a
Hernández-Bello Jorge a
Rosas-Rodríguez Jesús Alfredo b
Turrubiates-Hernández Francisco Javier a
García-Arellano Samuel a
González-Estevez Guillermo a
Ceja-Galvez Hazael Ramiro a
Oregon-Romero Edith a
López-Reyes Alberto c
https://orcid.org/0000-0002-2272-9260
Muñoz-Valle Jose Francisco a
a Instituto de Investigación en Ciencias Biomédicas (IICB), Centro Universitario de Ciencias de la Salud (CUCS), Universidad de Guadalajara (UdG) , Guadalajara, México
b Departamento de Ciencias Químico Biológicas y Agropecuarias, Universidad de Sonora Unidad Regional Sur , Navojoa, México
c Laboratorio de Gerociencias, Instituto Nacional de Rehabilitación “Luis Guillermo Ibarra Ibarra”, Secretaria de Salud , Ciudad de México, México
CONTACT Jose Francisco Muñoz-Valle biologiamolecular@hotmail.com; drjosefranciscomv@cucs.udg.mx Instituto de Investigación en Ciencias Biomédicas (IICB), Centro Universitario de Ciencias de la Salud (CUCS), Universidad de Guadalajara (UdG), Sierra Mojada 950, Independencia Oriente, Guadalajara, Jalisco 44340, México.
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© 2024 The Author(s). Published with license by Taylor & Francis Group, LLC.
2024
The Author(s)
https://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The terms on which this article has been published allow the posting of the Accepted Manuscript in a repository by the author(s) or with their consent.

ABSTRACT

To achieve global herd immunity, widespread vaccination is the most effective strategy. Vaccines stimulate the immune system, generating cytokines and chemokines, isotype antibodies, and neutralizing antibodies; all these molecules collectively provide a more comprehensive characterization of the immune response post-vaccination. We conducted a longitudinal study in northwestern Mexico, involving 120 individuals before vaccination and after the first dose of the SARS-CoV-2 vaccine, and 46 individuals after their second dose. Our findings reveal that antibody levels stabilize over time; cytokine levels generally increase following the first dose but decrease after the second dose and higher than normal levels in IgG1 and IgG3 concentrations are present. Most of the innate cytokines determined in this study were higher after the first dose of the vaccine. Regardless of previous infection history, this finding suggests that the first dose of the vaccine is crucial and may stimulate immunity by enhancing the innate immune response. Conversely, increased levels of IL-4, indicative of a Th2 response, were found in individuals without prior exposure to the virus and in those vaccinated with CoronaVac. These results suggest that the immune response to COVID-19 vaccines is multi-faceted, with preexisting immunity potentiating a more robust innate response. Vaccine type plays a critical role, with genetic vaccines favoring a Th1 response and inactivated vaccines like CoronaVac skewing toward a Th2 profile.

KEYWORDS

COVID-19
cytokines, neutralizing antibodies
SARS-CoV-2
SARS-CoV-2 vaccine
vaccination
National Council of Science and Technology A1-S-8774 University of Guadalajara This research was funded by the National Council of Science and Technology (CONACYT Ciencia Básica, Grant No. A1-S-8774 assigned to JFMV) and the University of Guadalajara through Strengthening Research and Postgraduate Studies 2022.
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pmcIntroduction

As of June 2023, Mexico has reported over 7.6 million COVID-19 cases and more than 334,336 deaths, according to data from the Open Data General Directorate of Epidemiology (https://datos.covid-19.conacyt.mx).

The most efficacious approach for achieving global herd immunity is through extensive vaccination. However, the emergence of SARS-CoV-2 variants has raised concerns about the potentially reduced effectiveness of COVID-19 vaccines.1,2 Therefore, developing effective vaccines against SARS-CoV-2 infection remains imperative for ensuring prolonged immunity and protection.

Various vaccine technologies have been developed, including inactivated virus vaccines, non-replicating viral vectors, and nucleic acid-based vaccines, such as DNA or mRNA vaccines.3 Inactivated virus vaccines are widely utilized and can expose the immune system to various viral proteins. An example of this type is CoronaVac, which has received emergency use authorization.4 This vaccine employs β-propiolactone inactivation of the virus, making it nonpathogenic, and, in this type of vaccine, the inclusion of adjuvants is required to elicit a robust immune response.5

Another innovative approach is the ChAdOx1 nCoV-19 vaccine, which uses a modified chimpanzee adenovirus (ChAdOx1) as a vector. The ChAdOx1 vector is engineered to carry genetic information encoding the full-length SARS-CoV-2 spike protein.6 In contrast, the BNT162b2 vaccine employs a different strategy, using mRNA extracted from the SARS-CoV-2 virus encapsulated within lipid nanoparticles. This vaccine introduces genetically modified RNA to trigger a controlled immune response.5

It has been reported that the time elapsed after vaccine administration affects its efficacy. For certain vaccines, significant efficacy is observed at least 14 days after the initial dose;7,8 however, other studies indicate that the effectiveness of single-dose vaccines could increase after 21 days.9,10 These data are important and contribute to the scientific evidence supporting vaccines. Nevertheless, these efficacy figures may not fully represent real-world effectiveness due to factors such as age distribution and other demographic variables. Therefore, studies like the present one are highly relevant.

Trained immunity, an innate memory response induced by infection or vaccination, is also pivotal in the body’s defense against potential infections.11 The transition from innate to adaptive immunity is vital for adequate protection against SARS-CoV-2.12 A key element of adaptative immunity is humoral immunity, which involves five classes of immunoglobulins: IgM, IgD, IgG, IgA, and IgE.13 These immunoglobulins are implicated in the humoral immune response against viral infections14 and possess various effector functions, including antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) by binding to C1q.15,16 Additionally, they can neutralize pathogens such as toxins and viruses.17

The primary target for COVID-19 vaccines is the SARS-CoV-2 spike glycoprotein, which is critical in eliciting neutralizing antibodies (NAbs). NAbs represent a crucial defense mechanism and highly indicate protection against the disease.18 All these vaccine platforms stimulate the immune system and produce molecules like cytokines and chemokines, which are vital for developing and maintaining innate immunity and adaptative immunity.19

Given that various factors, including genetics, vaccine platforms, cytokine response, and previous pathogen exposure (regardless of prior exposure to the SARS-CoV-2 virus) can influence vaccine efficacy,20–22 the primary objective of this study is to discern how individuals, both with and without prior SARS-CoV-2 infection, react to different vaccines. To achieve this, the study evaluates the levels of neutralizing antibodies against SARS-CoV-2, antibody isotypes IgM, IgA, IgG1, IgG2, IgG3, IgG4, and cytokine levels in the participants.

Materials and methods

Sample collection

This longitudinal study involved 120 individuals from northwest Mexico (Sonora, Mexico) 21 days after receiving the first COVID-19 vaccine, and 46 subjects assessed 21 days after their second dose.

Serum samples were collected before vaccination, with all samples being collected between July 2021 and August 2021 for various determinations. The study targeted young adults of both sexes, aged between 18 and 35 years old.

All participants provided informed consent statements upon enrollment at the University of Sonora, South Regional Unit, Sonora, Mexico. Additionally, clinical data regarding vaccination side effects and any prior SARS-CoV-2 infections were collected from each participant.

Individuals were classified into two groups based on their antibody test results and reported history of COVID-19 infection. The first group, labeled “with prior COVID-19,” included individuals who either tested positive for total and/or neutralizing antibodies or reported a prior COVID-19 infection. The second group, “without prior COVID-19,” consisted of those who tested negative for total and/or neutralizing antibodies. This classification was determined based on results obtained from lateral flow immunoassays and ELISA using the cPassTM SARS-CoV-2 Neutralizing Antibody Detection Kit.

The Wilcoxon Signed-Rank Test (matched pairs) was used for sample size calculation, comparing two repeated (paired) means in a single group, specifically comparing the first dose versus the second dose after each vaccine.n=Zα/2+Zβ2⋅2⋅s2d2

d=r1−r2

d=0.61−0.62

d=0.7

n=1.96+0.842⋅2⋅120.75

n=1.96+0.842⋅20.562

n=2.802⋅20.562

n=27.1

n=27 (for group)

n = sample size.

Zα/2 = Z-score (for α = 0.05, Zα/2 ≈ 1.96).

α = alpha error probability.

Zβ = Z-score (power of 0.80, Zβ≈0.84).

s2 = estimated variance of the differences.

d = effect size.

r = 0.6 (medium magnitude difference).

Determination of NAbs

The cPassTM SARS-CoV-2 neutralizing antibody detection kit (GenScript, Piscataway, NJ, USA) was used to identify neutralizing antibodies. NAbs were measured at three time points: before vaccination, 21 days after the first dose, and 21 days after the second dose of each vaccine. The timing for each variable measurement is specified in their respective sections.

The kit cutoff value is 30% signal inhibition. A value equal to or greater than 30% indicates a positive result and a value less than 30% indicates a negative result for neutralizing antibodies. The following formula is used to calculate the inhibition percent of the signal:% signal inhibition =1−OD450 sampleAverage OD450 negative control× 100%

Isotype antibodies determination

Total isotype antibodies (IgA, IgM, IgG1, IgG2, IgG3, and IgG4) were quantified using the Bio-Plex® MAGPIX™ system. This system employs fluorescently labeled magnetic beads, each with a unique spectral color code, allowing the discrimination of individual tests within a multiple suspension. These isotype levels were measured at two-time points: 21 days after the first dose and 21 days after the second dose of each vaccine. We employed a Bio-Rad catalog #171A3100M, and the manufacturer’s instructions were followed.

Serum cytokine levels determination

Cytokine levels were quantified using the Bio-Plex® MAGPIX™ system at three time points: before vaccination, 21 days after the first dose, and 21 days after the second dose of each vaccine. The manufacturer’s instructions were followed, and the fluorescence emitted from each was determined in the Bio-Plex MAGPIX reader (BIO-RAD). We used the Bio-Plex Pro Human Cytokine 27-Plex Panel (catalog #M500KCAF0Y). For this study, the following cytokines were considered: IFN-γ, TNF-α, IL-4, IL-1RA, IL-9, IL-8/CXCL8, IP-10/CXCL10, basic FGF, G-CSF, MCP-1/CCL2, MIP-1α/CCL3, MIP-1β/CCL4, RANTES/CCL5, eotaxin/CCL11, and PDGF-BB. The magnetic beads were read at 635 nm, and the reporter at 525 nm. The results were analyzed by adjusting the obtained values to the calibration curve.

Statistical analysis

The results were analyzed using RStudio V. 2022, SPSS V. 29, and GraphPad Prism 8.0 software. Continuous variables were represented as medians with interquartile range (IQR) for data description. The distribution of the data was assessed using the Kolmogorov-Smirnov test.

We applied the Mann-Whitney U test for comparisons between two independent groups, while the Wilcoxon rank test was employed for paired data analysis. The Friedman Test was used for dependent group comparisons, and the Kruskal-Walli’s test was used for independent groups. Chi-squared test was used for nominal variables. The Wilcoxon rank test and student t for dependent variables were used to calculate the difference between the two related tests. The correlations of cytokines were assessed using the Spearman correlation analysis.

Results

Eligibility and sampling

The process of selecting participants for the study began by recruiting 120 subjects from the general population – young adults, both with and without a history of SARS-CoV-2 infection, aged between 18 and 35 years old – who lived in southern Sonora (a northern state of Mexico) who expressed their willingness to participate and were considered eligible for the study, then, we proceeded with the follow-up assessments.

Of the initial cohort of 120 individuals who received the first vaccine dose, 74 participants were lost to follow-up, resulting in a final cohort of 46 individuals for the follow-up assessments, as shown in Figure 1. Therefore, the determinations were made based on the 46 individuals who completed the follow-up before and after vaccination. This final group comprised 28 individuals who received the CoronaVac vaccine, 7 who were administered the ChAdOx1 nCoV-19 vaccine, and 11 who received the BNT162b2 vaccine. It is important to emphasize that the variation in vaccine types administered was due to the availability of vaccines at the time of the study. The participants received the SARS-CoV-2 vaccine through random allocation, which was an integral part of the study design. Figure 1. Description of eligibility. 120 participants were recruited before vaccination and at the first dose, of which 74 were lost to follow-up; therefore, a total of 46 participants were the final sample for the second dose.

Side effects of SARS-CoV-2 vaccines after the first and second immunization

Adverse effects following vaccination can be widely influenced by factors such as age, sex, and the specific vaccine technology applied. Therefore, it was crucial to compare side effects across three vaccine groups, differentiated by the number of doses received (Table 1).Table 1. Side effects to CoronaVac, ChAdOx1 nCoV-19, and BNT162b2 after the first and second vaccination doses.

Side effects	First dose (n = 120)	Second dose (n = 46)	
CoronaVac (n = 73)	ChAdOx1 nCoV-19
(n = 7)	BNT162b2
(n = 40)	p-value	CoronaVac (n = 28)	ChAdOx1 nCoV-19
(n = 7)	BNT162b2
(n = 11)	p-value	
Any symptoms n(%)	22(30.1)	1(14.3)	26(65.0)	0.001	5(17.8)	1(14.3)	1(9.0)	0.204	
Headache n(%)	17(23.2)	5(71.4)	6(15.0)	0.005	11(39.2)	4(57.1)	0(0.0)	0.020	
Myalgia n(%)	37(50.7)	5(71.4)	8(20.0)	0.002	17(60.7)	5(71.4)	4(36.4)	0.266	
Fever n(%)	6(8.2)	1(14.3)	1(2.5)	0.358	6(21.4)	2(28.6)	4(36.4)	0.625	
Fatigue n(%)	3(4.1)	3(42.8)	1(2.5)	0.0001	4(14.2)	2(28.6)	0(0.0)	0.204	
Diarrhea n(%)	2(2.7)	0(0.0)	0(0.0)	0.520	1(3.6)	0(0.0)	0(0.0)	0.720	
Irritability n(%)	4(5.5)	1(14.3)	0(0.0)	0.146	5(17.9)	1(14.3)	0(0.0)	0.328	
Sore throat n(%)	3(4.1)	2(28.5)	0(0.0)	0.002	1(3.6)	0(0.0)	0(0.0)	0.720	
Rhinorrhea n(%)	4(5.5)	1(14.3)	0(0.0)	0.146	0(0.0)	1(14.3)	0(0.0)	0.058	
Excessive sleep n(%)	1(1.4)	0(0.0)	1(2.5)	0.849	4(14.3)	0(0.0)	1(9.1)	0.541	
Cough n(%)	1(1.4)	0(0.0)	0(0.0)	0.723	0(0.0)	0(0.0)	0(0.0)	–	
Arthralgia n(%)	0(0.0)	0(0.0)	0(0.0)	–	2(7.1)	1(14.3)	0(0.0)	0.478	
Conjunctivitis n(%)	0(0.0)	1(14.3)	0(0.0)	0.0001	0(0.0)	0(0.0)	0(0.0)	–	
Shaking chills n(%)	1(1.4)	0(0.0)	0(0.0)	0.723	2(7.1)	1(14.3)	0(0.0)	0.478	
Dizziness n(%)	0(0.0)	0(0.0)	1(2.5)	0.365	0(0.0)	0(0.0)	0(0.0)	–	
Application-site pain	0(0.0)	1(14.3)	0(0.0)	0.723	0(0.0)	2(28.6)	3(27.3)	0.541	
n(%)

The p-value was calculated using the Chi-squared test and for values >5 Cramer’s V to measure the strength of association.

Significant differences are shown in bold font (p < .05).

Individuals vaccinated with ChAdOx1 nCoV-19 showed a higher prevalence of side effects than those immunized with CoronaVac and BNT162b2. After the first dose, the most common symptoms were headache (23.33%), and myalgia (41.66%). After the second dose, headache remained prevalent among those vaccinated with ChAdOx1 nCoV-19 (57.1%) and CoronaVac (39.2%), but this was not observed in those vaccinated with BNT162b2.

A significant 85.7% of individuals vaccinated with ChAdOx1 nCoV-19 experienced adverse effects after the first dose. For BNT162b2, side effects such as myalgia (36.4%) and fever (36.4%) became most common after the second dose. Across all three vaccine types, there was a noticeable increase in adverse effects following the second dose compared to the first. While a higher proportion of individuals were asymptomatic in the first dose, those who did experience symptoms often reported more than one. In contrast, very few individuals remained asymptomatic after the second dose, and a significant proportion reported at least one adverse effect. Notably, symptoms like cough, dizziness, and arthralgia (the latter only after the first were not reported by any individual following the second dose).

Neutralizing antibodies in response to SARS-CoV-2 inactivated vaccine, mRNA, and adenovirus vaccine after the first and second doses

Figure 2 illustrates the levels of neutralizing antibodies elicited after the complete vaccination schedules for a) CoronaVac (n = 28), b) ChAdOx1 nCoV-19 (n = 7), and c) BNT162b2 (n = 11). Notably, after the first dose, the CoronaVac vaccine triggered a significant increase in antibody levels, showing a highly significant difference from the basal levels, with a p-value of <0.001. However, a slight decrease in neutralizing antibody levels was observed after administering the second dose of CoronaVac. This decrease did not reach statistical significance (p = .201). In contrast to CoronaVac, the individuals immunized with ChAdOx1 nCoV-19 and BNT162b2 vaccines consistently increased neutralizing antibody levels after each administered dose. Notably, statistically significant differences in antibody levels were observed following the vaccines’ first and second doses. Figure 2. Neutralizing antibodies were generated in response to CoronaVac, ChAdOx1 nCoV-19, and BNT162b2 vaccines. Differences were assessed using the Wilcoxon rank test. Results are presented in a table displaying medians 21 days after the first and the second vaccine doses. NAbs were measured 21 days after administering the first and second vaccine doses. The dotted line indicates the neutralization cutoff (>30%). n = 46.

In contrast to CoronaVac, the individuals immunized with ChAdOx1 nCoV-19 and BNT162b2 vaccines consistently increased neutralizing antibody levels after each administered dose. Notably, statistically significant differences in antibody levels were observed following the vaccines’ first and second doses.

Neutralizing antibodies in individuals with and without COVID-19 in both doses of SARS-CoV-2 vaccine

In both groups, an increase in the percentage of neutralization was observed after the administration of the first dose compared to basal levels (Figure 3). However, when comparing the first and second doses in the group with prior COVID-19 infection, no significant increase was observed between the two doses. Figure 3. Neutralizing antibodies in (a) individuals with COVID-19 (n = 26), and (b) individuals without COVID-19 (n = 20). Differences were assessed using the Wilcoxon rank test. The dotted line indicates the neutralization cutoff (>30%).

Isotype antibodies changes in vaccinated subjects from the first to the second dose of vaccination

After assessing the neutralizing capacity in individuals with prior SARS-CoV-2 infection, it became important to gain specific insights into the antibody isotypes involved in the immune response (Figure 4). Consequently, total antibody isotypes were analyzed in the serum of vaccinated individuals at 21 days post-administration of both the first and second vaccine doses. This comprehensive assessment encompassed IgM, IgA, and IgG antibodies and further examination of IgG subclasses, namely IgG1, IgG2, IgG3, and IgG4 (Figure 3). Figure 4. Total isotype antibody levels after the first and the second dose of the SARS-CoV-2 vaccine. The difference was calculated using the t-test or the Wilcoxon rank test for dependent samples. Results are presented in µg/ml for total isotype antibodies: (a) IgM, (b) IgA, (c) IgG1, (d) IgG2, (e) IgG3, (f) IgG4, 21 days after the first and the second dose of vaccination against SARS-CoV-2. The dotted line indicates the normal values.

The results revealed a significant decrease in IgA (p < .001) and IgG2 (p = .002) antibody levels following the administration of the second vaccine dose. However, it is noteworthy that both remained within the range of expected values.

Conversely, while no statistically significant differences were observed in the remaining antibody classes and subclasses, a discernible trend toward decreased levels was noted 21 days after the second vaccine dose. Additionally, levels of IgG1 and IgG3 were observed to surpass normal thresholds. As for IgM and IgG4, no statistically significant variations were detected, although it is worth mentioning that a few individuals exhibited levels above the established normal ranges in the case of IgG4.

Serum cytokine and chemokine levels in vaccinated subjects in the basal, first, and second doses of vaccination

In addition to eliciting the production of neutralizing antibodies, vaccination against SARS-CoV-2 prompts the secretion of cytokines and chemokines. A multiplex analysis was conducted to comprehensively assess the cytokine profiles after administering the first and second vaccine doses, focusing on a panel of cytokines of particular interest (Figure 5).

For most cytokines, including IFN-γ, TNFα, IL-1RA, IL-9, and chemokines such as IP-10/CXCL10, MIP-1β, MCP-1/CCL2, RANTES/CCL5 and PDGF-BB a consistent pattern emerged. Specifically, there was an increase in their levels after the first vaccine dose, followed by a subsequent decrease after the second dose.

However, the behavior of IL-4 diverged from the trend above. In this case, a decline in IL-4 levels was observed after the second dose, whereas no such decrease was noted following the first dose compared to basal levels. In contrast to these observations, an elevation in IL-8/CXCL8, C-GSF, and MIP-1α/CCL3 levels was detected after the second vaccine dose compared to basal levels. Figure 5. Comparison of cytokine and chemokine levels in basal levels and after the first and second dose of the SARS-CoV-2 vaccination. The cytokines and chemokines levels in serum before vaccination were compared 21 days after all vaccine technologies’ first and second doses. The difference was calculated using the Wilcoxon rank test for dependent samples. Data are provided in pg/ml. n=46.

Figure 5. (Continued).

Comparison of serum cytokines and chemokines levels in basal levels, first and second dose of vaccination divided by individuals with and without prior COVID-19

After assessing the cytokine and chemokine levels following the administering of both the initial and booster doses of the SARS-CoV-2 vaccine in the complete study population, a comparative analysis was essential. This analysis stratified individuals into two subgroups: those who had previously experienced a COVID-19 infection (n = 26) and those who had not encountered such an infection (n = 20). Subsequently, serum cytokines and chemokines were analyzed for all three subject groups who had completed the vaccination regimen.

For most cytokines and chemokines, including IFN-γ, TNF-α, IL-1RA, IL-9, IL-8/CXCL8, IP-10/CXCL10, MCP-1/CCL2, MIP-1β/CCL4, RANTES/CCL5 and PDGF-BB a similar pattern was observed in both the groups (with and without prior COVID-19 history). Specifically, these cytokines and chemokines exhibited higher median levels after the first vaccine dose, followed by a subsequent decrease in levels following the second dose (Table 2).Table 2. Comparison of serum cytokine levels in basal, first, and second doses in individuals with or without prior COVID-19.

Cytokine/Chemokine	With prior COVID-19	Without prior COVID-19	
Basal n = 261	First dose n = 261	Second dose n = 261	p-value2	Basal n = 201	First dose n = 201	Second dose n = 201	p-value2	
IFN-γ	1.93 (1.63–2.86)	3.64 (3.11–4.17)	1.63 (1.63–2.62)	0.001	2.94 (1.63–3.68)	3.53 (2.72–4.18)	1.63 (1.63–2.12)	0.001	
TNF-α	29.22 (20.19–35.82)	36.39 (29.24–43.97)	10.15 (5.19–19.81)	0.001	20.25 (23.94–37.29)	36.75 (29.49–39.71)	7.37 (5.19–11.32)	0.001	
IL-4	1.93 (1.48–2.35)	1.48 (1.16–2.35)	1.53 (1.28–2.04)	0.023	1.97 (1.62–2.25)	2.26 (1.93–3.43)	1.53 (1.17–1.69)	0.02	
IL-1RA	103.09 (66.39–124.57)	181.77 (146.03–316.19)	100.77 (55.09–176.10)	0.001	119.76 (67.07–144.89)	184 (141–251.07)	84.8 (58.72–117.01)	0.001	
IL-9	72.58 (67.13–80.49)	94.92 (83.88–107.98)	36.67 (31.71–43.61)	0.001	72.31 (62.74–78.27)	87.13 (74.48–94.14)	30.86 (26.56–40.81)	0.001	
IL-8/CXCL8	4.1 (3.25–5.93)	6.8 (4.52–23.28)	4.1 (4.1–12.33)	0.007	4.1 (4–4.8)	4.49 (3.5–12.97)	4.1 (4.1–13.27)	0.139	
IP-10/CXCL10	168.03 (131–216)	253.55 (159.78–338.25)	55.96 (47.40–79.03)	0.001	184.62 (138.60–252.04)	208.36 (155.22–222.13)	64.96 (42.36–77.2)	0.001	
Basic FGF	4.97 (3.36–6.49)	5.7 (4.61–8.66)	4.67 (3.89–11.33)	0.123	4.66 (4.06–5.98)	5.32(4.48 – 6.38)	5.24 (3.36–8.25)	0.468	
G-CSF	77.06 (51.08–129.64)	97.34 (54.71–222.58)	86.96 (40.81–416.96)	0.607	78.72 (57.63–99.18)	93.51 (48.82–152.86)	107.34 (46.28–329.60)	0.287	
MCP-1/CCL2	16.33 (9.68–22.96)	28.53 (21.54–39.17)	7.25 (2.72–11.09)	0.001	16.69 (11.23–23)	24.37 (17–97–33.68)	9.91 (3.95–10.69)	0.001	
MIP-1α/CCL3	1.20 (0.82–1.72)	1.52 (1.24–2.97)	1.07 (0.72–7.15)	0.542	1.24 (0.88–1.63)	1.42 (.077–2.44)	1.29 (0.81–5.2)	0.035	
MIP-1β/CCL4	69.49 (66.81–81.57)	93.92 (85.95–108.80)	36.91 (29.62–53.05)	0.001	67.73 (60.29–76.47)	86.33 (79.99–96.57)	36.89 (22.76–42.47)	0.001	
RANTES/CCL5	1320.43 (1024.53–1630.92)	2386.18 (1917.49–2770.42)	720.78 (485.01–1021.55)	0.001	1261.88 (1005.82–1715.32)	2037.63 (1769.16–2577.55)	525.47 (384.98–822.87)	0.001	
Eotaxin/CCL11	79.53 (62.77–119.90)	50.78 (40.70–74.76)	50.41(26.13 -78.95)	0.002	73.82 (52.86–84.35)	67.60 (29.32–89.45)	28.54 (5.5–47.42)	0.008	
PDGF-BB	288.06 (143.94–414.92)	1596.75 (694.01–2671.47)	250.91 (79.41–337.02)	0.001	287.39 (197.07–333.15)	1436.03 (731.12–1776.88)	322.97 (137.26–491.31)	0.001	
Median (Q1-Q3).

Friedman Test.

Significant differences are shown in bold font (p<.05).

In contrast, no significant differences were detected in the levels of Basic FGF, G-CSF, and MIP-1α/CCL3 between the groups of individuals with a history of prior COVID-19 infection. Notably, the concentration of IL-4 increased after the initial vaccine dose and decreased upon administering the second dose in individuals without prior COVID-19 infection. However, these concentrations exhibited a notable decrease in individuals with prior infection, with lower levels persisting after the second vaccine dose.

Comparative analysis of serum cytokine and chemokine levels in individuals with and without previous COVID-19 infection, segregated based on their first and second vaccine doses

To validate the preceding findings, examining whether distinctions existed between individuals with a history of COVID-19 infection and those without was imperative. Consequently, a comparative analysis was conducted, stratifying individuals into two groups: those with prior COVID-19 infection (n = 26) and those without (n = 20), differentiated by the administration of both the first and second doses of all administered vaccines (Table 3).Table 3. Serum cytokine levels in individuals with or without prior COVID-19 after the first and second doses.

Cytokine/Chemokine	First dose	Second dose	
With prior COVID-19	Without prior COVID-19	p-value2	With prior COVID-19	Without prior COVID-19	p-value2	
IFN-γ	3.64 (3.11–4.17)	3.53 (2.84–4.07)	0.485	1.63 (1.63–2.62)	1.63 (1.63–2.12)	0.287	
TNF-α	36.39 (29.24–43.78)	36.75 (29.49–39.71)	0.465	10.16 (5.19–19.71)	7.37 (5.19–11.17)	0.179	
IL-4	1.48 (1.60–2.35)	2.26 (1.93–3.43)	0.009	1.53 (1.28–2.04)	1.53.3 (1.17–1.69)	0.340	
IL-1RA	181.78 (148.52–303.75)	184 (142.44–250.87)	0.634	100.77 (55.78–172.37)	84.80 (61.67–116.90)	0.393	
IL-9	94.93 (84.17–107.80)	87.13 (75.44–93.85)	0.030	36.68 (31.92–43.24)	30.86 (26.67–40.62)	0.116	
IL-8/CXCL8	6.80 (4.67–21.31)	4.49 (3.71–12.75)	0.140	4.10 (4.10–11.87)	4.10 (4.10–12.61)	0.579	
IP-10/CXCL10	253.56 (163.60–330.88)	208.36 (155.86–221.94)	0.126	55.96 (50.48–78.85)	64.97 (42.71–77.20)	0.965	
Basic FGF	5.70 (4.70–8.64)	5.32 (35.48–89.40)	0.346	4.67 (4.05–11.32)	5.24 (3.38–8.16)	0.730	
G-CSF	97.34 (56.91–221.06)	93.51 (49.68–148.16)	0.400	86.97 (41.39–408.62)	107.35 (49.06–327.90)	0.825	
MCP-1/CCL2	28.53 (21.78–38.10)	24.38 (18.24–33.52)	0.287	7.25 (2.77–10.93)	7.92 (4.06–10.67)	0.965	
MIP-1α/CCL3	1.53 (1.28–2.97)	1.42 (0.78–2.38)	0.176	1.07 (0.73–7.10)	1.29 (0.83–5.13)	0.698	
MIP-1β/CCL4	93.91 (86.04–106.72)	86.33 (80.02–96.11)	0.044	36.92 (29.64–52.74)	36.89 (23.07–42.29)	0.258	
RANTES/CCL5	2386.18 (1818.24–2716.22)	2037.63 (1801.95–2559.36)	0.595	720.78 (516.17–1014.04)	525.48 (385.88–819.18)	0.191	
Eotaxin/CCL11	50.78 (41.60–72.50)	67.60 (35.48–89.40)	0.375	50.42 (26.64–77.74)	28.54 (6.02–45.47)	0.007	
PDGF-BB	1596.76 (731.26–2444.76)	1436.03 (734.87–1769.30)	0.341	250.92 (86.85–328.58)	322.97 (190.23–478.27)	0.059	
Median (Q1-Q3).

Mann-Whitney’s U Test.

Significant differences are shown in bold font (p<.05).

Indeed, following the first vaccine dose, in individuals without a history of prior COVID-19 infection, most cytokines exhibited no statistically significant disparities between the two groups. However, notable distinctions were observed in the levels of IL-9 and MIP-1β/CCL4. Specifically, the levels of these three cytokines were notably elevated in individuals who had previously experienced the disease. Conversely, IL-4 showed increased levels of individuals without a history of COVID-19 after the first dose.

Concerning the second vaccine dose, no differences were observed for IFN-γ in the levels of this cytokine in individuals with or without a prior COVID-19 history. A comparable trend was noted for Eotaxin as well. However, PDGF-BB exhibited distinct results, with higher levels observed in individuals who had not previously contracted the disease.

Comparison of serum cytokine and chemokines levels between individuals who received CoronaVac, ChAdOx1 nCoV-19, and BNT162b2 vaccines after the first and second immunization

Once identifying distinct cytokine patterns, we embarked on an analysis focused on the specific vaccines administered (Table 3). At the 21-day mark following the first vaccine dose, we observed elevated levels of TNF-α in the group vaccinated with ChAdOx1 nCoV-19. We heightened levels of RANTES/CCL5 (p = .029) and IP-10 (p = .051) in the group vaccinated with CoronaVac compared to individuals who received other vaccine formulations.

Conversely, at the 21-day interval after the second vaccine dose, a significant elevation in cytokines/chemokines, including IFN-γ, IL-1RA, IL-8/CXCL8, G-CSF, MIP-1α/CCL3 and RANTES/CCL5 was noted in the group vaccinated with BNT162b2, compared to those who received CoronaVac and ChAdOx1 nCoV-19 vaccines, with statistically significant differences. In contrast, individuals vaccinated with CoronaVac exhibited a significant increase in IL-4 (p = .049) after the second dose compared to other groups (Table 4).Table 4. Comparison between serum cytokine levels in CoronaVac, ChAdOx1 nCoV-19, and BNT162b2 after the first and second doses.

Cytokine/Chemokine	First dose	Second dose	
CoronaVac n=281	ChAdOx1 nCoV-19 n=71	BNT162b2 n=111	p-value2	CoronaVac n=281	ChAdOx1 nCoV-19 n=71	BNT162b2 n=111	p-value2	
IFN-γ	3.48 (2.65–4.37)	4.02 (3.8–4.88)	3.6 (2.08–4.21)	0.207	1.63 (1.63–2.03)	1.63 (1.63–2.14)	3.17 (1.88–3.17)	0.005	
TNF-α	34.61 (27.52–40.61)	40.96 (37.96–43.78)	37.62 (34.78–44.57)	0.051	9.19 (5.19–16.27)	6.55 (5.19–11.47)	9.38 (6.28–19.71)	0.648	
IL-4	2.12 (1.21–2.72)	2.07 (2.22–2.95)	1.65 (1.16–2.02)	0.214	1.68 (1.34–2.04)	1.28 (1.16–1.4)	1.45 (1.16–2.4)	0.049	
IL-1RA	185.21 (141.83–291.38)	186.1 (84.49–251.27)	156.54 (145.27–270.3)	0.784	83.98 (62.10–148.40)	55.78 (47.37–85.11)	156.54 (99.53–187.32)	0.042	
IL-9	96.52 (82.67–108.34)	87.69 (81.63–101.85)	85.69 (77.34–94.01)	0.150	35.07 (27.23–42.34)	30.53 (21.85–35.41)	35.62 (27.28–49.76)	0.313	
IL-8/CXCL8	6.33 (3.92–21.02)	13.19 (4.66–22.06)	4.1 (4.1–8.01)	0.240	4.10 (2.44–4.10)	4.1 (4.1–21.97)	16.75 (5.85–23.02)	0.001	
IP-10/CXCL10	276.53 (184.44–364.27)	213.19 (168.14–235.76)	157.12 (97.8–194.89)	0.006	55.96 (42.99–79.00)	70.35 (45.82–77.2)	56.63 (38.17–106.65)	0.992	
Basic FGF	5.78 (4.81–7.22)	6.14 (5.24–7.05)	5.32 (3.36–5.92)	0.175	4.94 (3.42–6.81)	3.42 (3.36–6.56)	7.98 (4.05–12.92)	0.111	
G-CSF	93.41 (51.12–213.84)	157.57
(70.08 – 292.39)	82.03 (47.97–104.55)	0.206	66.59 (44.34–230.92)	54.76 (33.73–324.49)	442 (201.65–781.12)	0.027	
MCP-1/CCL2	27.96 (20.97–36.72)	24.05 (21.66–31.19)	28.32 (15.17–42.43)	0.827	7.33 (4.39–10.69)	4.83 (2.27–8.61)	8.99 (2.27–21.66)	0.390	
MIP-1α/CCL3	1.51 (0.99–2.90)	2.51 (0.92–4.27)	1.38 (0.96–1.57)	0.159	0.96 (0.74–2.01)	0.84 (0.55–4.99)	7.9 (1.75–9.31)	0.009	
MIP-1β/CCL4	91.93 (82.31–108.63)	86.53 (80.53–97.04)	87.91 (85.55–98.35)	0.827	36.17 (29.58–42.58)	28.72 (22.46–41.90)	54 (32.09–66.32)	0.067	
RANTES/CCL5	2455.39 (1970.49–3268.65)	2134.44 (1867.53–2595.75)	1818.24 (1349.33–2143.67)	0.029	623.05 (437.15–944.52)	391.23 (384.09–568.30)	836.69 (729.74–1072.18)	0.049	
Eotaxin/CCL11	48.76(33.24 - 84.06)	69.13 (52.56–99.97)	56.88 (41.6–89.28)	0.175	4.71 (24.50–68.88)	19.05 (3.65–31.74)	52.17 (8.24–89.45)	0.058	
PDGF-BB	1652.93 (976.55–2355.60)	1409.57 (703.66–1784.48)	742.37 (328.02–2116.22)	0.297	295.62 (77.64–439.23)	287.98 (49.14–317.14)	311.51 (193.02-
464.5)	0.368	
Median (Q1-Q3).

2Krurskal Wallis Test.

Significant differences are shown in bold font (p<.05).

Irrespective of the vaccine type administered, more cytokine changes were observed after the second dose, with medians generally lower than those after the first. An exception to this trend was observed in the case of IL-8, where medians were higher in individuals who received the second vaccine dose.

Correlation of cytokines

Cytokines play a fundamental role in developing and maintaining adaptive immunity in response to infection and vaccination. Therefore, we performed a correlation matrix analysis between the determined cytokines and chemokines (Figure 6). Figure 6. Correlation of cytokines and chemokines levels before and after the first and second vaccination dose. The correlation of cytokines and chemokines levels was performed in serum (a) before vaccination, (b) 21 days after the first dose, and (c) the second dose of all vaccine technologies. Spearman test was used for the analysis. X= non-significant difference (p<.05).

We observed a major correlation between Basic FGF and IL-4 (p = .71) with other cytokines in basal levels, and after the first dose, these correlations disappeared. However, in the first dose, we noticed a correlation between IFN-γ and IL-1RA (p = .57). We detected a correlation between IL-9 with G-CSF, IL-8, MIP-1β/CCL4, PDGF, and RANTES/CCL5.

On the other hand, after 21 days of the second dose of the vaccine, we perceived more significant correlations between cytokines. The correlations between IFN-γ and IL-1RA (p = .73), IP-10 (p = .5), and RANTES/CCL5 (p = .55), and IFN-γ also correlated with IL-8 and MIP-1β/CCL4.

Discussion

The global mass vaccination effort against SARS-CoV-2 has unveiled significant heterogeneity in immune responses, underscoring the need for a detailed understanding of these variations. This study aimed to dissect these responses, focusing on cytokine patterns, antibody responses, and side effects post-vaccination with different vaccine types. Our results show a heterogeneity of self-reported side effects in response to vaccination and revealed that adverse effects were more pronounced after the first vaccine dose than the second one.

Individuals who received the ChAdOx1 nCoV-19 vaccine displayed more side effects than the other two, which could be attributed to the complexity of adenovirus vector vaccines, which replicate in mammalian cell cultures and can elicit heterogeneous immune responses, potentially leading to an increased occurrence of adverse effects.23 Furthermore, these findings align with the results obtained in our research group, where a higher incidence of adverse effects was observed among individuals who received the non-replicating viral vector Ad5-nCoV vaccine.24 They describe that the increased reactogenicity may stem from the interaction between the viral vector and the vaccine’s antigenic component.24

Our study, consistent with previous research in the Mexican population,25 indicates a substantial increase in NAbs post-first vaccine dose. This supports that in individuals with prior COVID-19, a single dose of the SARS-CoV-2 vaccine elicits a NAbs response similar to what was observed in individuals without previous SARS-CoV-2 infection who received two doses of the vaccine.22,26 This is significant in the context of limited vaccine supplies and could provide valuable insights for future vaccine distribution strategies.

Samples were collected 21 days after each dose, as it can take up to 21 days for antibodies against the virus to develop. Furthermore, following the second vaccine dose, a decline in NAbs was observed in some individuals. Consequently, we performed a stratified analysis based on the administered vaccine. The results indicated that individuals exhibiting reduced NAbs predominantly belonged to the CoronaVac vaccine group. These findings align with reports in the Mexican population, where a more robust response was noted after the first dose application compared to the second dose of the SARS-CoV-2 vaccine.25 However, the decline in NAbs following the second dose in some individuals, particularly among CoronaVac recipients, raises questions about the duration of immunity and the potential need for booster doses. Vaccine effectiveness was minimal for the first two weeks post-initial dose, but it rose significantly in the third and fourth weeks before the second dose. Therefore, it was important to make the comparison after three weeks of each dose to observe the differences.10 Samples were collected 21 days after each dose because it can take up to 21 days for antibodies against the virus to develop. Additionally, the highest peak of the antibodies could be reflected at this time.27

Some changes were observed when determining the NAbs after the second dose of the vaccine was administered. Individuals with prior COVID-19 did not exhibit an additional increase in NAbs after the second dose compared to the first one. This observation aligns with findings reported by Goel et al., who noted that individuals who had recovered from SARS-CoV-2 exhibited an expansion of S-specific memory B cells and RBD after the initial immunization. However, they did not observe an additional increase after the second vaccine dose.28 This could be due to the stabilization of antibodies over time, resulting in a less pronounced boost post-second dose.

Considering that most NAbs against SARS-CoV-2 primarily target the Receptor-Binding Domain (RBD) – approximately 90% of them – the S protein´s conformation is crucial.29 Different conformations of the S protein, pre- or post-fusion with the host membrane, have been reported, making a pre-fusion optimal for generating NAbs.23 mRNA vaccines include two mutations that have been incorporated to stabilize the S2 subunit of the S protein,30 preventing the conformational change toward the prefusion structure. In contrast, the ChAdOx1 nCoV-19 vaccine, despite lacking these mutations, still retains a prefusion conformation similar to the mRNA vaccine.23 However, a study focusing on the S protein in inactivated virus vaccines has revealed that SARS-CoV-2 viruses treated with β-propiolactone exhibit a predominance of post-fusion conformation.31 This observation may potentially explain the results obtained in our study regarding the lower production of NAbs in response to the CoronaVac vaccine.

In addition to assessing neutralizing capacity following prior SARS-CoV-2 infection, we also investigate the antibody isotypes participating in the immune response after the first and second vaccine doses.

A decrease in the concentrations of IgA and IgG2 was observed after the second dose; however, these levels remained within the normal ranges. This observation coincides with a study by Campos-Mata and collaborators, which proposes that the above could reflect two phases of humoral immunity. The initial phase is characterized by a rapid release of minimally mutated classes and subclasses of antibodies via the extrafollicular pathway, possibly produced by short-lived plasma cells. The subsequent phase involves the emergence of IgG produced by long-lived plasma cells, potentially originating from germinal centers.32

In the context of SARS-CoV-2 infection, a predominant IgG1 and IgG3 antibody response has been documented.33,34 These two subclasses of IgG antibodies may have viral neutralization capacity, opsonization, and the ability to activate the complement system in viral respiratory infections.35 Supporting this, our findings indicated increased IgG1 and IgG3 levels above average values, although no statistically significant differences were observed between groups. These results are consistent with studies reported by Qi et al., suggesting that these antibodies can exhibit relative stability over several months, with IgG being the most resilient.36 Regarding IgG4, a few individuals exhibited levels above the normal range. It could be attributed to the fact that IgG4 primarily responds to a different class of infections, specifically extracellular parasites.37

Conversely, no significant differences were noticed in the IgM results.

It is well-established that IgM antibodies are the first immunoglobulin species involved in the primary immune response.38,39 However, these antibodies may also persist in the serum of individuals vaccinated against COVID-19, as observed in studies of those infected with the virus.40,41 It is crucial to conduct further research to analyze the kinetics of these antibodies over time in vaccinated individuals.

Consequently, it can be inferred that IgM levels stabilize over time, much like IgG1 and IgG3. Therefore, it is imperative to explore specific immunoglobulins for SARS-CoV-2 shortly.

On the other hand, numerous prior studies have examined serum cytokine and chemokine levels and their association with COVID-19 severity,42–44 and relatively few have explored these markers in response to vaccination. Cytokines can play several functional roles in different immune contexts; each could be classified or belong to more than one component.45 In that sense, it was crucial to investigate the cytokines and chemokines induced by vaccination to identify markers facilitating the development of protective immunity against SARS-CoV-2.

Our results indicated a general drop in immune responses after the second vaccine dose, regardless of the vaccine type. Most cytokines exhibited an increase following the first dose and a subsequent decrease after the second dose. A study conducted by Vega-Magaña and collaborators46 measured the response of T cells and observed a decline over time, suggesting this might be attributed to persistent antigen stimulation, leading to T cell depletion and subsequently reduced cytokine production.

Although the CoronaVac vaccine did not induce the highest antibody responses, elevated levels of IL-4 were observed in individuals vaccinated with CoronaVac. IL-4 is likely more involved in humoral immune responses than other cytokines, as it coincided with antibody stabilization over time. Additionally, a tendency toward higher IL-4 levels was observed in individuals without preexisting immunity or prior COVID-19, possibly indicating its role in stimulating B cell development and antibody production.45

Elevated chemokine RANTES/CCL5 levels were also observed following the first vaccine dose, irrespective of the administered vaccine. However, higher levels were noted in the CoronaVac group when analyzed by vaccine type. A study conducted by Zhao et al. in individuals with mild, moderate, and severe SARS-CoV-2 infections reported increased RANTES/CCL5 levels in mild cases, correlating with lymphocyte counts and suggesting a protective role of lymphocytes in the disease.47 This observation aligns with the previously mentioned findings, where individuals immunized with CoronaVac exhibited elevated IL-4 levels compared to those who received genetic vaccines. Consequently, one could hypothesize that the induction of the innate immune response after the initial dose is irrespective of the specific vaccine administered. Conversely, a heightened humoral immune response is discerned in individuals inoculated with CoronaVac following the second dose, contrasting with responses elicited by other vaccine modalities.

In contrast, most individuals vaccinated with the BNT162b2 vaccine had prior COVID-19 and elevated levels of IFN-γ were observed in individuals vaccinated with the BNT162b2 mRNA vaccine and an elevated median in individuals with a history of COVID-19. Administration of the BNT162b2 mRNA vaccine also induced higher levels of other cytokines/chemokines, such as IL-1RA, RANTES/CCL5, IL-8, G-CSF, and MIP-1α, which play crucial roles in innate immune responses,48 adaptive immunity development, and immunological memory. Moreover, the analysis revealed a Th1 cytokine response (IFN-γ) in BNT162b2-vaccinated individuals, but a low percentage of IL-4 expression associated with the Th2 response, highlighting the importance of booster vaccinations for preventing SARS-CoV-2 reinfection.

IL-1RA, a cytokine with an anti-inflammatory effect, plays a significant role in immune response modulation. Zhao et al. suggested that in mild infections, the adaptive responses of the virus can cancel the activity of IL-1RA, whereas in severe cases, elevated cytokine levels were found.47 This suggests that IL-1RA may help prevent patients from developing severe disease.47 In our study, we found higher levels of IL-1RA in the second dose of the BNT162b2 vaccinated group; however, we also observed increases in proinflammatory cytokines (IFN-γ, RANTES, IL-8, C-GSF, and MIP-1α/CCL3) in the same group. This suggests a potential compensatory mechanism in play, balancing proinflammatory and anti-inflammatory responses.

Furthermore, our study observed higher levels of TNF-α and a tendency for elevated IFN-γ levels in individuals vaccinated with the first dose of ChAdOx1 nCoV-19. These findings align with a study by Jiang et al., which reported that higher IFN and pro-inflammatory cytokine responses, including IFNs and TNF-α, were induced in human cells (PBMCs) vaccinated with ChAdOx1 nCoV-19 compared to mRNA vaccines.49 This difference could be attributed to genetic vaccines’ ability to trigger interferon production by recognizing innate sensors essential for initiating inflammatory responses.50

Finally, correlations were identified between IFN-γ and IL-1RA, as well as IFN-γ and IP-10. Other studies also noted that these correlations have been associated with higher antibody titers.51,52 Additionally, IP-10/CXCL10 is known to influence the differentiation of activated B cells into plasma cells.53

In our study, potential limitations should be considered when interpreting the results: first, the possibility that our sample size may be insufficiently representative. This limitation could impact the generalizability of our findings, particularly in a study exploring complex immunological responses to vaccines. Second, the loss of participants during follow-up is a well-documented issue in longitudinal research. We hypothesize that this may be due to a lack of commitment to follow-up, insufficient information about the research in certain regions, and logistical challenges hindering participation in scientific studies. Additionally, some participants were unavailable during the follow-up period, and others did not complete the survey. These factors may affect the external validity of the results due to the impact on the sample size. These limitations underscore the need to explore other aspects of the immune response and to consider the reasons for the missed follow-up visits and the time interval between infection and vaccination, which could provide valuable insights for future research. Although previous studies have examined objectives similar to those of the present research; most focus on only one or two types of vaccines. A key strength of the current study is its examination of three types of vaccines, which were critical in addressing the pandemic, assessing participants before and after vaccination.

Participants were encouraged to take part in the research, with an explanation that the results would contribute to scientific advancements. They were given flexibility regarding the time and day of their participation. Additionally, they were offered the option to complete the survey online if they had time constraints. They also received regular reminders for their appointments and were assured of confidentiality.

Another limitation of this study is the possibility that the timing of our cytokine measurements, precisely at 21 days post-vaccination, may not align with the peak activity of specific cytokines. Given that some cytokines have a short half-life or rapid response time, our measurements might not fully capture the initial or peak response dynamics. For that reason, further research is required to investigate how cytokines and chemokines correlate with cellular responses. However, this information is essential to illuminate the interplay between innate and adaptive immune responses. Nevertheless, this study provides a comprehensive overview of the immune response dynamics following homologous vaccination employing three distinct vaccine technologies targeting SARS-CoV-2.

Conclusions

Most of the innate cytokines determined in this study were higher after the first dose of the vaccine. Regardless of previous infection history, this finding suggests that the first dose of the vaccine is crucial and may stimulate immunity by enhancing the innate immune response. In contrast, individuals without prior exposure to the virus and lacking neutralizing antibodies exhibit a delayed innate immune response. In these individuals, the induction of cytokines, particularly IL-4, associated with a Th2 response, appears crucial. This cytokine activity is likely important for promoting B cell differentiation and the subsequent production of neutralizing antibodies, essential for effective protection against the virus.

José Francisco Muñoz-Valle is a doctor in Molecular Biology in Medicine from the University of Guadalajara (UdeG), with a postdoctorate at the University of Oxford, United Kingdom (2005). Founder of the Biomedical Sciences Research Institute. Rector of the University Center for Health Sciences (2019-2022 and 2022-2025) of the UdeG. Member of the SNII level 3, with more than three hundred publications and more than five thousand three hundred citations. He has trained human resources at different academic levels. Recipient of one hundred national and international awards, highlighting the 2014 Research Award from the Mexican Academy of Sciences. Member of various academies and general coordinator of the COVID-19 Health Situation Room of the UdeG. Representative of Mexico before the International Federation of Clinical Chemistry.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Author contributions

Conceptualization, J.F.M-V.; methodology, D.L.P.B. and F.J.T.H.; software, D.L.P.B. and F.J.T.H.; validation, J.F.M-V., J.H.B.; formal analysis, D.L.P.B., H.R.C.G.; investigation, D.L.P.B.; resources, J.F.M-V.; data curation, D.L.P.B., M.G.M.F., J.A.R.R.; writing – original draft preparation, D.L.P.B.; writing – review and editing, J.F.M-V., J.H.B, A.G.L.R., E.O.R., G.G.E.; visualization, J.F.M-V.; supervision, J.F.M-V. and S.G.A.; project administration, J.F.M-V.; funding acquisition, J.F.M-V. All authors have read and agreed to the published version of the manuscript.

Data availability statement

The data from this study are available from the corresponding author upon reasonable request.

Informed consent statement

Informed consent was obtained from all subjects involved in the study.
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