
==== Front
J Prim Care Community Health
J Prim Care Community Health
JPC
spjpc
Journal of Primary Care & Community Health
2150-1319
2150-1327
SAGE Publications Sage CA: Los Angeles, CA

39257165
10.1177/21501319241273213
10.1177_21501319241273213
Original Research Article
Association Between SARS-CoV-2 Vaccination and Development of Antinuclear Antibodies Among Students
Laskowski Karol 1*
Paszkiewicz Justyna 1*
Plewik Dorota 1
Szepeluk Adam 1
https://orcid.org/0000-0001-8606-2509
Hozyasz Kamil K. 1
1 John Paul II University in Biała Podlaska, Poland
Kamil K Hozyasz, Department of Nursing, Faculty of Health Sciences, John Paul II University in Biała Podlaska, 95/97 Sidorska Street, Biała Podlaska, 21-500 Poland. Email: khozyasz@gmail.com
* These authors contributed equally to this work.

10 9 2024
Jan-Dec 2024
15 2150131924127321319 5 2024
20 6 2024
12 7 2024
© The Author(s) 2024
2024
SAGE Publications Inc unless otherwise noted. Manuscript content on this site is licensed under Creative Commons Licenses
https://creativecommons.org/licenses/by-nc/4.0/ This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (https://creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages (https://us.sagepub.com/en-us/nam/open-access-at-sage).
Introduction:

Antinuclear antibodies (ANA) are the hallmark of many connective tissue disorders (including lupus), which comprise roughly 5% to 10% of chronic debilitating diseases causing morbidity and mortality. In society, fear of these diseases increases illness-related uncertainty because the prognosis of progression is often difficult to determine and at least some symptoms fluctuate unpredictably. In the anti-vaccination movement, the question of the possible connection between vaccination and connective tissue disorders and other autoimmune diseases has grown to rank as an important argument for rejecting vaccination. In 2021, every fifth Polish first-degree nursing student decided to not be vaccinated against COVID-19.

Objective:

This study aimed to explore the prevalence of antinuclear antibodies in students vaccinated and unvaccinated against COVID-19.

Participants and Methods:

A single university cross-sectional study was performed in a small academic centre in Poland, where 210 students were recruited in 2022. All the participants were screened for SARS-CoV-2 IgG and antinuclear antibodies.

Results:

The mean age of the students who rejected vaccination was higher than that of those who were vaccinated. Among nursing students, 30.0% of vaccinated and 58.3% of unvaccinated individuals had COVID-19. The frequency of antinuclear antibodies was 3 times lower in vaccinated students than in unvaccinated students (2/159 vs 2/51; P > .05).

Conclusion:

The results of our study did not confirm the rationality of rejecting vaccinations against COVID-19 for fear of developing autoimmune diseases among healthy students.

COVID-19 vaccination
anti-nuclear antibodies
nursing students
autoimmune diseases
Państwowa Szkoła Wyższa im. Papieża Jana Pawła II w Białej Podlaskiej https://doi.org/10.13039/501100014420 PB/4/22 cover-dateJanuary-December 2024
typesetterts1
==== Body
pmcIntroduction

Connective tissue disorders (CTD), commonly referred to as systemic autoimmune diseases or collagenopathies, comprise roughly 5% to 10% of chronic debilitating diseases that cause morbidity and mortality.1-3 CTD is a wide variety of related diseases characterised by dysregulation of immune homeostasis which enables the activation of immune cells to attack autoantigens, resulting in chronic increased inflammation and multi-tissue damage. Detailed characteristics of more than a hundred CTDs have already been described. The clinical hallmarks of CTD overlap with those of diseases caused by pathogenic microorganisms and other nonimmune-mediated disorders. Patients may experience considerable illness-related uncertainty because the prognosis of disease progression is often difficult to determine and at least some symptoms fluctuate unpredictably.1-3

As these diseases are autoimmune in nature, a plethora of well-described autoantibodies are useful for differential diagnosis. Since their first description, antinuclear antibodies (ANA) have been the hallmarks of many CTD (so-called ANA-associated rheumatic diseases), such as Sjögren’s syndrome, systemic lupus erythematosus (SLE), mixed connective tissue disease, idiopathic inflammatory myopathies, and systemic sclerosis. ANA is also important in the diagnosis of autoimmune hepatitis and primary biliary cholangitis. Trace concentrations of ANA are measurable in approximately 10% to 30% of healthy individuals. 3 Indirect immunofluorescence microscopy (IIF) using human epithelial HEp-2 cell substrates is considered the most accurate method for ANA detection in serum samples. 3 However, many commercial enzymes immunoassay kits with variable performance are available for routine ANA screening.

Vaccines may initiate autoimmunity through mechanisms such as molecular mimicry, T-cell activation without antigen recognition, epitope spreading, and an anti-idiotypic network. However, there are no elaborated criteria for the diagnosis of vaccine-induced CTD, which has to be evaluated on a case-by-case basis. 4 The question of the connection between vaccination and CTD and other autoimmune diseases has been discussed in the scientific literature for decades, 5 and in the anti-vaccination movement, it has grown to the rank of an important argument for rejecting vaccination. 6

It was expected that students, particularly biomedical students, would not be susceptible to anti-vaccine rhetoric during the pandemic.7,8 However additionally, during the COVID-19 pandemic, part of the political scene in Poland, including the parliament, was openly against restrictions and presented vaccinations against SARS-CoV-2 as a threat to people, especially in pre- and reproductive age. 9 In 2021, over 22% of 793 first-degree nursing students from 12 Polish universities were not vaccinated against COVID-19, and every second participant declared that they would stick to their choice. 10 In a study conducted at the turn of December 2023 and January 2024 almost 20% of medical students from Poznań, one of the largest and most economically developed cities in Poland, believe at least 1 anti-vax conspiracy theory. 8 One of the arguments for not being vaccinated against COVID-19, even by medical students or healthcare workers, was the fear of developing autoimmune diseases.8,10,11 The dissemination of information about vaccinations inconsistent with research results on social media may be one of the most important factors in the denial of vaccination against COVID-19. 12 A recent study by Bartosiewicz et al 13 showed that sources of knowledge about vaccines influenced attitudes toward vaccination and were the main predictors of fear of the pandemic among Polish biomedical students. The absorption and processing of scientific information and news-sharing processes in social media are subjects of intensive research. 14 Appropriate emotional content can increase the willingness of social media users to share medical findings from scientific papers on vaccination and infectious diseases. 15

Thus, (I) the purpose of the study was to compare the prevalence of ANA in vaccinated against COVID-19 and unvaccinated healthy individuals, and (II) our study focused on students, with the assumption that the results obtained, apart from the expected scientific value, will inspire the students themselves and for their social media activity.

Materials and Methods

Study Design, Sample, and Setting

This single-university cross-sectional study was performed in a small academic center, educating approximately 3000 students in Biała Podlaska, Poland. The vast majority of vaccinated students received the BNT162b2 mRNA COVID-19 vaccine intramuscularly (Comirnaty, Pfizer, Inc., and BioNTech). The active substance of the preparation is the mRNA that encodes the spike protein of SARS-CoV-2 and acts as an antigen. Of the approximately 2500 students actively participating in the classes, it was intended to recruit 10%.

Basic data, data on vaccination against COVID-19, the course of COVID-19, and blood samples were collected from participants between April 2022 and December 2022.

Inclusion/Exclusion criteria

The inclusion criteria were a negative history of immunological diseases, vaccination with the Comirnaty vaccine (vaccinated group of students), and no anti-COVID-19 vaccinations (unvaccinated group of students). 2. The exclusion criteria were age <18 years, foreign students, pregnancy, and use of drugs that affect the immune system (eg, anabolic glucocorticoids, cytostatics, tacrolimus, and azithromycin).

Research Question

Are there differences in the prevalence of ANA among students vaccinated and unvaccinated against COVID-19?

Ethical Considerations

The study protocol was approved by the local ethics committee (approval number: 10/2021). The authors assert that all procedures contributing to this study are compatible with the ethical standards of the relevant national and institutional committees on human experimentation and the 1975 Helsinki Declaration, as revised in 2008. Written informed consent was obtained from all the participants. The members of the research team invited all healthy students with no history of immunological disturbances to participate in the study and those who agreed to complete the questionnaire.

Before conducting the research, the participants were introduced to the purpose of the research and their rights and obligations. They were also informed of their voluntary participation, anonymity, and ability to leave the study at any time.

Laboratory Tests

Blood samples were immediately transferred to the laboratory, where IgG antibody titres to the SARS-CoV-2 S1-protein were determined using an Anti-SARS-CoV-2 QuantiVac ELISA IgG assay (Euroimmune, Germany). The ELISA was developed on 96-well microplates coated with the SARS-CoV-2 S1 domain, which was expressed recombinantly in human embryonic kidney 293 cells. Quantification of S1-specific IgG was performed according to the manufacturer’s instructions using a 6-point calibration curve covering a range of 1 to 120 relative units (RU)/ mL. Serum samples yielding results in the analytical range were each time retested at a higher dilution. In each test run, positive and negative controls were included. Seropositivity was defined as >11 RU/mL. Serologic screening for antinuclear antibodies (ANA 1) was performed using the ANA Screen 11 ELISA IgG (Euroimmune, Germany) with a cutoff antibody index >1.45 for seropositivity. Autoantibodies in ANA 1—positive participants were confirmed by indirect immunofluorescence (IIF) on Hep-2 cells (ANA 2; Euroimmune, Germany) and by line immunoblotting (ANA 3 Profile with 15 antigens; Euroimmune, Germany) on a Euroblot Master 44 device using Euroline Scan Software (Euroimmune, Germany). The IIF cutoff was set at 1:160 dilution, which was in accordance with clinical practice. 3

Data Management and Analysis

Collected data were verified for completeness, quality, and consistency. All the data collected on the paper were transcribed into electronic databases for analysis. None of the participants dropped out of this study. Statistical analysis was performed using the Polish version of TIBCO Software Inc., Palo Alto, Santa Clara, CA, USA (2017). The compliance of the data with the normal distribution was assessed using the Shapiro-Wilk test (P > .05 for normal distribution). Categorical data were compared using the chi-squared or Fisher’s exact test based on the size of the smallest subgroup. The comparison of proportions between groups was performed using the Mann-Whitney U test. All qualitative findings are presented as n (%). Statistical significance was set at P < .05.

Results

Sample Characteristics

This study analyzed the data of 210 students, of whom 159 were vaccinated against COVID-19 and 51 were unvaccinated. The most numerous participants were nursing and emergency medical students, constituting 69.2% and 10.1% of the vaccinated group and 47.1% and 3.9% of the unvaccinated group, respectively. A comparison between the 2 groups in terms of demographics and history of COVID-19 is shown in Table 1. The age of students who rejected the offer of vaccination was statistically significantly higher than that of those who were vaccinated. More than one-third of the unvaccinated students were male, with 22% of the vaccinated students being male. Almost half of the unvaccinated students reported COVID-19 to less than 1 in 3 vaccinated. Among the nursing students, 30% of the vaccinated (33/110) and 58.3% of the unvaccinated (14/24) had COVID-19. Loss of taste and smell during the illness was reported by 70.8% of unvaccinated students (17/24) and 78.3% of vaccinated students (36/46), but a high fever above 39°C was observed in 18.5% of unvaccinated students (5/24) and only 1 vaccinated student (2.2%).

Table 1. Baselin Characteristics of the Study Population.

	Unvaccinated students (n = 51)	Vaccinated students (n = 159)	P	
Age, years mean ± SD	29.9 ± 12.2	23.6 ± 5.0	.002	
Gender M/F, n/n	18/33	35/124	.057	
Faculty	
 Nursing, n (%)	24 (47.1)	110 (69.2)	<.001	
 Paramedics, n (%)	2 (3.9)	16 (10.1)	
 Dietetics, n (%)	0	6 (4.0)	
 Others*, n (%)	25 (49.0)	27 (17.0)	
History of COVID-19, n	24	46	.017	
The length of time between getting sick from COVID-19 and blood drawn, months, mean ± SD	No data	9.3 ± 6.5		
The length of time between the last vaccination and blood drawn, months, mean ± SD	Not applicable	4.4 ± 3.5		
Number of doses of the vaccine	
 First	Not applicable	56		
 Second	Not applicable	62		
 Third	Not applicable	41		
ANA positivity, n	2	2	.249	
* i.a. Finance and accounting, Management, Agriculture, Pedagogy.

Anti-SARS-CoV antibodies

No antibodies against SARS-CoV-2 were detected in any of the fourth unvaccinated students (Table 2). Only 1 vaccinated student did not develop any antibodies. High titres of antibodies above 2000 RU/mL were not found in unvaccinated students. There were no statistically significant differences in the levels of antibodies against SARS-CoV-2 in the unvaccinated students who thought they had not been infected with the virus and those who were sick, which was confirmed by real-time polymerase chain reaction (RT-PCR) test, which is considered the gold standard for viral detection, 16 (Table 3).

Table 2. Comparison of Anti-SARS-CoV-2 IgG Antibody Levels in Unvaccinated and Vaccinated Students.

Groups of students	Anti-SARS-CoV-2 IgG concentrations (RU/mL)	
<11 (negative)	11-100	101-500	501-1000	1001-2000	>2000	
n	%	n	%	n	%	n	%	N	%	n	%	
Unvaccinated (n = 51)	14	27.5	20	39.2	14	27.5	2	3.9	1	2	0	0	
Vaccinated	Total (n = 159)	1	0.6	8	5.0	62	39.0	49	31.0	26	16.4	13	8.2	
No pre-vaccination history of COVID-19 (n = 131)	0	0	6	4.6	50	38.2	44	33.6	21	16.0	10	7.6	
pre-vaccination history of COVID-19 (n = 28)	1	3.6	2	7.1	12	42.9	5	17.9	5	17.9	3	10.7	
P < .001.

Table 3. Comparison of Anti-SARS-CoV-2 IgG Antibody Levels in Unvaccinated Students Without Prior COVID-19 and Convalescents With SARS-CoV-2 Infection Confirmed With RT PCR Test.

Groups of unvaccinated students	Anti-SARS-CoV-2 IgG concentrations (RU/mL)	
<11 (negative)	11-100	101-500	501-1000	1001-2000	>2000	
n	%	n	%	n	%	n	%	n	%	n	%	
Without prior COVID-19 (n = 27)	8	29.6	10	37.0	9	33.3	0	0	0	0	0	0	
Convalescents (PCR +; n = 18)	3	16.7	8	40.4	4	22.2	2	11.1	1	5.5	0	0	
P = .210.

ANA assessments

In the groups of vaccinated and unvaccinated students, 2 cases with ANA 1 were found and confirmed by IIF (1.3% vs 3.9%, P = .25). Both unvaccinated ANA-positive female students had COVID-19 and developed anti-SARS-CoV-2 antibodies (Table 4). Vaccinated female students with ANA had no COVID-19 history and titters of anti-SARS-CoV-2 antibodies below 2000 RU/mL. In 1 ANA-positive unvaccinated student, we found increased anti-Ro-52 antibodies (Table 5). Notably, no signs of new-onset inflammatory or autoimmune disease were reported by the students, regardless of ANA development.

Table 4. Characteristics of ANA-Positive Students. a

Vaccination status	Age (years)	Gender	ANA ratio	Anti-SARS-CoV-2 IgG concentration (RU/mL)	History of COVID-19	The length of time between the last dose of vaccine and blood drawn (months)	Number of doses of the vaccine	
Unvaccinated	
 Case 1	37	Female	2.44	410	Yes b	Not applicable	0	
 Case 2	39	Female	6.87	43	Yes c	Not applicable	0	
Vaccinated	
 Case 3	27	Female	1.61	569	Not	1	3	
 Case 4	21	Female	1.46	1454	Not	10	2	
a All ANA-positive students studied health care related fields. More detailed data was not provided for reasons of anonymity.

b Illness unconfirmed by RT PCR test, loss of smell and taste persisted for 5 days, there was no high fever.

c Illness unconfirmed by RT PCR test, there was no loss of smell and taste and no high fever.

Table 5. Results of ANA-positive Sera Analysis Based on Indirect Immunofluorescence on Hep-2 Cells (ANA2) and Line Immunoblot (ANA3).

Methods of analysis	Unvaccinated students	Vaccinated students	
Case 1	Case 2	Case 3	Case 4	
ANA2	
 Granular fluorescence pattern	1:320	1:320	1:160	1:160	
 Cytoplasmic fluorescence pattern	1:160	Negative	Negative	1:160	
ANA3	
 Immunoblot results	
  Anti-RNP	7.0	1.0	2.0	0	
  Anti-Sm	2.0	0	2.0	0	
  Anti-SSA	1.0	2.0	1.0	0	
  Anti-Ro-52	2.0	178.0	2.0	3.0	
  Anti-SSB	3.0	5.0	2.0	1.0	
  Anti-Scl-70	13.0	2.0	1.0	1.0	
  Anti-PM-Scl	2.0	2.0	3.0	34.0	
  Anti-Jo-1	2.0	3.0	3.0	3.0	
  Anti-CB	1.0	1.0	1.0	2.0	
  Anti-PCNA	2.0	2.0	1.0	3.0	
  Anti-dsDNA	5.0	0	6.0	3.0	
  Anti-NUC	2.0	0	1.0	1.0	
  Anti-HI	1.0	1.0	0	0	
  Anti-RIB	2.0	0	1.0	1.0	
  Anti-M2	1.0	1	2.0	1.0	
  Control	112.0	137	132.0	121.0	

Discussion

The influential voice of the worldwide “anti-vax” movement led many people to postpone or even reject the anti-COVID vaccination in general and therefore jeopardised the efforts to slow down the pandemic. 11 In our study, ANA was 3 times less common in vaccinated students than in unvaccinated students, and 1 unvaccinated ANA-positive student developed anti-Ro-52 antibodies. To the best of our knowledge, this is the first study to compare the ANA levels between vaccinated against COVID-19 and unvaccinated healthy students. The 52 kDa protein Ro-52 (TRIM21) is mainly expressed in lymphoid tissues and serves as a shared target for autoantibodies in collagenopaties. 17 Convincing proof for the pathogenic role of anti-Ro-52 antibodies in systemic autoimmune diseases was provided by transfer experiments of anti-Ro-52 sera into New Zealand mixed mice and the onset of disease. 18

The quantitative determination of anti-SARS-CoV-2 antibodies is pivotal for estimating the humoral response to infection caused by this virus and vaccination. In this study, we used the Anti-SARS-CoV-2 Quantivac ELISA IgG assay kit which targets the S1 surface protein. This test kit had a high sensitivity for detecting anti-SARS-CoV-2 antibodies at a rate of 97.8%. 19 Three of every 4 unvaccinated students developed anti-SARS-CoV-2 antibodies. Seven out of 10 unvaccinated students who denied suffering from COVID-19 had anti-SARS-CoV-2 antibodies, which is consistent with observations about frequent clinically asymptomatic cases of COVID-19. 20 The longer the pandemic lasts, the more the interview about not getting sick from COVID-19 should be treated as not the lack of illness, but the avoidance of more severe disease. Interestingly, no antibodies were detected in the 3 unvaccinated students whose SARS-CoV-2 infection was confirmed by positive PCR test results.

SARS-CoV-2 has had a devastating impact on human health and the world in many other dimensions, such as the global economy and politics, leading to the rapid development of vaccines to prevent the spread of the disease. Currently, the vaccine is widely available worldwide and has noticeably lowered the spread of SARS-CoV-2. The pioneering use of mRNA vaccines has been accompanied by concerns regarding potential autoimmunization.4,21 The relationship between vaccination against COVID-19 and the occurrence of autoimmune disorders has been documented in numerous clinical case reports.22,23 The largest cohort to date of 11 patients with new-onset autoimmune diseases with positive ANA related to the mRNA-based COVID-19 vaccine from a single center study (Khobar, Saudi Arabia) included 4 SLE patients (22, 25, 33, and 37 years-old), 2 patients with inflammatory arthritis (26 and 27 years-old), and 1 patient each with Sjögren’s syndrome (44 years-old), dermatomyositis (46 years-old), autoimmune hepatitis (16 years-old), autoimmune pancreatitis (21 years-old), and antisynthetase syndrome (18 years-old). 24 Very recent database search in PubMed, EMBASE, Web of Science, and Scopus identified 16 new-onset cases of SLE (44% under the age of 30 years) following COVID-19 vaccination reported between 2021 and 2023 worldwide, 25 which does not change the fact that, in statistical terms, this is a marginal phenomenon. 21 Most cases were associated with mRNA-based vaccines (82.5%) and the first dose (56.2%). 25 The time elapsed from vaccine administration to the onset of symptoms of SLE was between 1 and 30 days (average 11.8 ± 8.1 days). Two-thirds of the patients who developed the disease had no family members or were immunologically predisposed. Studies based on case reports are particularly vulnerable to risks of over-interpretation and selection bias. Nevertheless, the South Korean cohort of 9.2 million individuals who had received at least 1 dose of the mRNA-based COVID-19 vaccine and had no prior diagnosis of COVID-19 in comparison to a historical control cohort was at considerably higher risk of developing SLE (adjusted hazard ratio = 1.53; 95% CI = 1.09-2.13). 26

Autoimmune disease experts are consistently convinced that in the general population, the benefits of vaccination against COVID-19 significantly outweigh the possible risks and should motivate people to be vaccinated.4,21 Even for patients with existing CTD at risk of flares with vaccination, in a “living document” the American College of Rheumatology recommends vaccination because the risk of poor outcomes from COVID-19 infection outweighs the risk of increased disease activity. 27 Several studies have reported that autoantibodies are prevalent in COVID-19 patients. COVID-19 patients who tested positive for ANA tended to have a severe condition and worse prognosis, including the prediction of a 28-day mortality period in critically ill patients. 28 Persistently positive ANA 1-year post-COVID-19 is associated with persistent symptoms and inflammation, similar to the so-called long COVID for a subset of COVID-19 survivors. 29 COVID-19 is a predisposing factor for auto-reactivity and is implicated in pathways that contribute to the onset of autoimmunity and may result in autoimmune diseases such as CTD, autoimmune hemolytic anemia, and multiple sclerosis. 30

Limitations and Recommendations

This study has some limitations. Recruitment plans were implemented by 84 % of participants. Signing up to participate in our study required blood sampling, which probably had a demotivating effect, probably less in the group of biomedical students. 31 Interestingly, in the recent study investigating vaccine hesitancy without planned blood collection only 17.7% of Poznań’s medical students decided to participate by completing an online survey. 8 The number of unvaccinated participants included in the study was low, making it difficult to draw definite conclusions regarding the development of ANA in this subgroup. Moreover, when discussing the frequency of ANA-positive results in the study group, we must emphasise that we did not know the frequency of their occurrence before the COVID-19 pandemic. Both contracting COVID-19 and vaccination against it can initiate a cascade of processes leading to the production of ANA.3,21,24,30 In the unvaccinated group, vaccination was certainly not a risk factor for developing ANA. In turn, in the group of vaccinated students, even if they do not contract COVID-19, exposure to SARS-CoV-2 is a potential risk factor for the development of ANA due to the high frequency of asymptomatic SARS-CoV-2 infections, especially among young people. In our study, the majority of the unvaccinated students with no history of COVID-19 disease developed anti-SARS-CoV-2 IgG antibodies.

The average age differed significantly statistically in the groups of vaccinated and unvaccinated students, but this can be directly explained by the specificity of the latter group and not faulty recruitment. 8 The study is also limited by the anti-SARS-CoV-2 antibody test, which was designed to detect post-vaccination antibodies and, to a lesser extent, post-infection antibodies. 32 Furthermore, some cross-reactivity between SARS-CoV-2-specific antibodies and other endemic RNA viruses, causing respiratory tract infections, has been reported and can reduce test credibility 33 ; however, the data sheet does not account for cross-reactivity with samples from patients infected with SARS-CoV-1, HCoV-OC43, or HCoV-229E (Euroimmun. Anti-SARS-CoV-2 Quantivac IgG ELISA. Package Insert). Extended serological diagnostics could enable a more complete understanding of the immune status of unvaccinated students who have had COVID-19 and do not have anti-SARS-CoV-2 antibodies as well as that of students who did not demonstrate a post-vaccination humoral response or demonstrated a modest response.

Referring the presented results to other population groups limits the specificity of the study group, which are students below 30 years of age on average.

Implications for Practice

Our study provides important information that could be valuable for healthy individuals who have difficulty deciding whether to get vaccinated because of the fear of developing autoimmune diseases. The data from our study could be used to inform communities that do not support or trust vaccination on the lack of the likelihood of auto-immunoreactivity associated with CTD after vaccination against COVID-19. Moreover, medical professionals dealing with vaccinations in the student population should direct a particularly reinforced message to older and male students, who raise concerns. Male students seem to be more susceptible to the message of anti-vaccination movements. 8

Conclusions

We did not find a presumed more frequent occurrence of ANA in healthy individuals vaccinated against COVID-19 among a group of 210 students from a small university located in the rural part of Poland. The results of our study did not confirm the rationality of rejecting vaccinations against COVID-19 for fear of developing autoimmune diseases among healthy students. It is important to emphasise the need for further studies on the connection between vaccination and autoimmune diseases.

The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Funding: The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This study was funded by the Akademia Bialska Center of Excellence under Grants No PB/3/2021 and PB/4/2022.

Ethical Approval: Permission to perform this research was obtained from the local ethics committee (Komisja Bioetyczna przy Akademii Bialskiej im. Jana Pawła II, no. 10/2021). Before conducting the research, participants were introduced to the purpose of the research and their rights and obligations. They were also informed of their voluntary participation, anonymity, and ability to leave at any time.

ORCID iD: Kamil K. Hozyasz https://orcid.org/0000-0001-8606-2509

Data Availability Statement: The datasets analyzed during the current study are available from the corresponding author on reasonable request.
==== Refs
References

1 Simons G Falahee M. Fear of the unknown: can we help individuals with a systemic autoimmune rheumatic disease deal with uncertainty?. J Rhematol. 2022;49 (9 ):977-979.
2 Conrad N Misra S Verbakel JY , et al . Incidence, prevalence, and co-occurrence of autoimmune disorders over time and by age, sex, and socioeconomic status: a population based cohort study of 22 million individuals in UK. Lancet. 2023; 401 (10291 ):1878-1890.37156255
3 Bonroy C Vercammen M Fierz W , et al . Detection of antinuclear antibodies: recommendations from EFLM, EASI and ICAP. Clin Chem Lab Med. 2023;61 (7 ):1167-1198.36989417
4 Chen Y Xu Z Wang P Li X-M Shuai Z-W Ye D-Q Pan H-F. New-onset autoimmune phenomena post-COVID-19 vaccination. Immunology. 2022;165 :386-401.34957554
5 Shoenfeld Y Aron-Maor A. Vaccination and autoimmunity – ‘vaccinosis’: a dangerous liaison?. J Autoimmun. 2000;14 (1 ):1-10.10648110
6 Poland GA Jacobson RM. The clinician’s guide to the anti-vaccinationists’ galaxy. Hum Immunol. 2012;73 (8 ): 859-866.22504410
7 Szmyd B Bartoszek A Karuga FF Staniecka K Błaszczyk M Radek M. Medical students and SARS-CoV-2 vaccination: attitude and behaviors. Vaccines. 2021;9 :128.33562872
8 Domaradzki J Jabkowski P Walkowiak D. Investigating beliefs in anti-vax conspiracy theories among medical students. Vaccines. 2024;12 :359.38675741
9 Turska-Kawa A Pilch I. Political beliefs and the acceptance of the SARS-CoV-2 pandemic restrictions. The case of Poland. PLoS One. 2022;17 (3 ):e0264502.
10 Gotlib J Sobierajski T Jaworski M , et al . Vaccinate, do not hesitate!. Vaccination readiness against COVID-19 among Polish nursing undergraduate students: a national cross-sectional survey. Vaccines. 2021;9 :1029.34579265
11 Holzmann-Littig C Frank T Schmaderer C , et al . COVID-19 vaccines: fear of side effects among German health care workers. Vaccines. 2022;10 (5 ):689.35632445
12 Wawrzuta D Jaworski M Gotlib J Panczyk M. Characteristics of antivaccine messages on social media: systematic review. J Med Internet Res. 2021;23 (6 ):e24564.
13 Bartosiewicz A Łuszczki E Bartosiewicz A Dereń K Oleksy Ł Stolarczyk A. COVID-19-related predictors of fear and attitude to vaccination displayed by Polish students. Vaccines. 2022;10 :1524.36146600
14 Cascini F Pantovic A Al-Ajlouni YA , et al . Social media and attitudes towards a COVID-19 vaccination: a systematic review of the literature. eClinicalMedicine. 2022;48 :101454.35611343
15 Wawrzuta D Jaworski M Gotlib J Panczyk M. Social media sharing of articles about measles in a European context: text analysis study. J Med Internet Res. 2021;23 (11 ):e30150.
16 Velavan TP Meyer CG. COVID-19: a PCR-defined pandemic. Int J Infect Dis. 2021;103 :278-279.33271291
17 Lee AY Lin M-W Reed JH. Anti-Ro52/TRIM21 serological subsets identify differential clinical and laboratory parameters. Clin Rheumatol. 2022;41 :3495-3501.35871174
18 Szczerba BM Kaplonek P Wolska N , et al . Interaction between innate immunity and Ro52-induced antibody causes Sjögren’s syndrome-like disorder in mice. Ann Rheum Dis. 2016;75 :617-622.25906316
19 Patel EU Bloch EM Clarke W , et al . Comparative performance of five commercially available serologic assays to detect antibodies to SARS-CoV-2 and identify individuals with high neutralizing titers. J Clin Microbiol. 2021;59 (2 ):e02257-20.
20 Syangtan G Bista S Dawadi P , et al . Asymptomatic SARS-CoV-2 carriers: a systematic review and meta-analysis. Front Public Health. 2021;8 :587374.33553089
21 Guo M Liu X Chen X Li Q. Insights into new-onset autoimmune diseases after COVID-19 vaccination. Autoimmun Rev. 2023;22 (7 ):103340.37075917
22 Liu J Wu H Xia S-L. New-onset arthritis following COVID-19 vaccination: a systemic review of case reports. Vaccines. 2023;11 :665.36992249
23 Gips JR Woreta TA. It can’t be a coincidence: a comparison of cases of autoimmune hepatitis after vaccination against COVID-19. ACG Case Rep J. 2023;10 :e00695.
24 Alqatari S Ismail M Hasan M , et al . Emergence of post COVID-19 vaccine autoimmune diseases: a single center study. Infect Drug Resist. 2023;16 :1263-1278.36910517
25 Cahuapaza-Gutierrez NL. Systemic lupus erythematosus following COVID-19 vaccination. A systematic review of case reports and case series. Lupus. 2024;33 (4 ):375-386.38315894
26 Lee S Jung S-W Jeon JJ Choe SJ. Long-term risk of autoimmune diseases after mRNA-based COVID-19 vaccination: a Korean population-based nationwide cohort study. Res Square. Published online April 17, 2024. doi:10.21203/rs.3.rs-4254839/v1
27 Curtis JR Johnson SR Anthony DD , et al . Amercican Collage of Rheumatology Guidance for COVID-19 vaccination in patients with rheumatic and musculoskeletal diseases: version 5. Arthritis Rheumatol. 2023;75 (1 ):E1-E16.
28 Park SH Suh JW Yang K-S , et al . Clinical significance of antinuclear antibody positivity in patients with severe Coronavirus disease 2019. J Korean Intern Med. 2023,38 (3 ):417-426.
29 Son K Jamil R Chowdhury A , et al . Circulating anti-nuclear autoantibodies in COVID-19 survivors predict long COVID symptoms. Eur Respir J. 2023;61 (1 ):2200970.36137590
30 Zebardast A Hasanzadeh A Ebrahimian Shiadeh SA Tourani M Yahyapour Y. COVID-19: a trigger of autoimmune diseases. Cell Biol Int. 2023;47 :848-858.36740221
31 Postema A Ferreira JA van der Klis F de Melker H Mollema L. Investigating sources of non-response bias in a population-based seroprevalence study of vaccine-preventable diseases in the Netherlands. BMC Infect Dis. 2024;24 (1 ):249.38395775
32 Tretyn A Szczepanek J Skorupa M , et al . Differences in the concentration of anti-SARS-CoV-2 IgG antibodies post-COVID-19 recovery or post vaccination. Cells. 2021;10 :1952.34440721
33 Nagappa B Marimuthu Y. Seroconversion rate and diagnostic accuracy of serological tests for Coronavirus 2019. Clin Infect Dis. 2020;71 (11 ):3016.
