The relationship between disgust sensitivity and serum cytokine levels in non-pregnant women: a generalization of previous results on pregnant population

The relationship between disgust sensitivity and serum cytokine levels in non-pregnant women: a generalization of previous results on pregnant population | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js'; var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s); })(); Skip to main content Preprints Collections Wiley Open Research IET Open Research Ecological Society of Japan All Collections About About Authorea FAQs Contact Us Quick Search anywhere Search for preprint articles, keywords, etc. Search Search ADVANCED SEARCH SCROLL This is a preprint and has not been peer reviewed. Data may be preliminary. 19 August 2025 V1 Latest version Share on The relationship between disgust sensitivity and serum cytokine levels in non-pregnant women: a generalization of previous results on pregnant population Authors : Daniela Dlouhá 0000-0002-2968-1992 [email protected] , Josef Včelák , and Šárka Kaňková Authors Info & Affiliations https://doi.org/10.22541/au.175558739.94517887/v1 244 views 92 downloads Contents Abstract Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract The behavioural immune system functions via the emotion of disgust, which triggers avoidant reactions that protect the individual from pathogens. The behavioural immune system is closely linked with physiological immune system, their mutually compensatory and preparatory functions are discussed. A recent study on pregnant women supported the notion that disgust increases when the physiological immune system is not sufficiently activated. The aim of this current study was to confirm these results on 78 healthy non-pregnant women, measuring immune activity via multiplex immunoassay and disgust sensitivity via the Disgust Scale Revised (DS-R), Three Domains of Disgust Scale (TDDS) and Tactile Disgust Stimuli Set, as well as Perceived Vulnerability to Disease. As in the previous study, we showed a significant negative association between disgust and certain pro-inflammatory cytokines, specifically between DS-R core disgust and MCP-1 and DS-R contamination disgust and IL-1β and TNF-α, suggesting a compensatory relationship between the systems. Simultaneously, we observed positive associations between IL-5 and IL-12 and both the DS-R core and overall scores, as well as between the TDDS pathogen domain and MIP-1β. These associations may reflect some psychological states and traits, as elevated levels of IL-5, IL-12 and MIP-1β have been linked to neuroticism, depression, and disorders accompanied by anxiety. We found no significant correlations between cytokine levels and both tactile disgust and perceived vulnerability to disease. Our findings thus support both the adaptive function of disgust and its links to mental health and point to the complexity of the behavioural immune system. The relationship between disgust sensitivity and serum cytokine levels in non-pregnant women: a generalization of previous results on pregnant population Daniela Dlouhá 1 , Josef Včelák 2 , Šárka Kaňková 1 1 Department of Philosophy and History of Science, Faculty of Science, Charles University, Viničná 7, 128 44 Prague 2, Czech Republic 2 Department of Molecular Endocrinology, Institute of Endocrinology, Národní 8, 116 94 Prague 1, Czech Republic Corresponding author: Daniela Dlouhá Tel.: +420 721840938 E-mail: [email protected] ORCID: 0000-0002-2968-1992 Department of Philosophy and History of Science, Faculty of Science, Charles University, Prague, Viničná 7, 128 44, Czech Republic Co-author contact information: Šárka Kaňková – [email protected] Josef Včelák – [email protected] Abstract The behavioural immune system functions via the emotion of disgust, which triggers avoidant reactions that protect the individual from pathogens. The behavioural immune system is closely linked with physiological immune system, their mutually compensatory and preparatory functions are discussed. A recent study on pregnant women supported the notion that disgust increases when the physiological immune system is not sufficiently activated. The aim of this current study was to confirm these results on 78 healthy non-pregnant women, measuring immune activity via multiplex immunoassay and disgust sensitivity via the Disgust Scale Revised (DS-R), Three Domains of Disgust Scale (TDDS) and Tactile Disgust Stimuli Set, as well as Perceived Vulnerability to Disease. As in the previous study, we showed a significant negative association between disgust and certain pro-inflammatory cytokines, specifically between DS-R core disgust and MCP-1 and DS-R contamination disgust and IL-1β and TNF-α, suggesting a compensatory relationship between the systems. Simultaneously, we observed positive associations between IL-5 and IL-12 and both the DS-R core and overall scores, as well as between the TDDS pathogen domain and MIP-1β. These associations may reflect some psychological states and traits, as elevated levels of IL-5, IL-12 and MIP-1β have been linked to neuroticism, depression, and disorders accompanied by anxiety. We found no significant correlations between cytokine levels and both tactile disgust and perceived vulnerability to disease. Our findings thus support both the adaptive function of disgust and its links to mental health and point to the complexity of the behavioural immune system. Keywords: Immune markers, disgust, pathogen avoidance, behavioural immune system, physiological immune system, mental health Introduction The body’s physiological immune system (PIS) is a complex network of organs, cells and proteins that can facilitate (amongst others) an immune reaction, which protects the body against pathogens. However, this activation of the PIS has a number of drawbacks; it is quite energetically costly and the individual is weakened during the course of the reaction. Because it is activated after the individual comes into contact with the pathogen, it would seem beneficial if the contact could in some way be prevented and the system would not have to activate as often. That is the role of the behavioural immune system (BIS) [1] and its affective component - the emotion of disgust. A sign of an activated BIS is that we feel disgusted upon seeing stimuli that can pose a pathogen threat. The disgust reaction then in turn facilitates a behavioural reaction – avoidance of the stimuli and of the pathogen threat. The activity of these two systems seems to be closely integrated, though the exact relationship is not yet clear. One such integration is the hypothesis that when the immune system is suppressed or less active, the activity of the behavioural immune system should be elevated to compensate. This premise is also known as the compensatory prophylaxis hypothesis [2]. Several studies have focused on testing this hypothesis. Some showed that disgust was elevated in people who reported being more frequently sick [3], recently sick [4], more vulnerable to disease [5], lived in an environment with higher risk of infection [6–8] or faced a pandemic threat [9]. A study also showed that recently ill people reacted more negatively to faces displaying disease cues [10], but it has not been successfully replicated [11]. Additionally, a number of other studies did not show support for aspects of this hypothesis, specifically that frequency of illness in childhood didn’t impact disgust sensitivity [12] and neither did the use of immunosuppressive medication, which even had an opposite effect [13]. While these studies tested the relationship between the activity of the BIS and the PIS, the indicators of immune system activity were only based on self-reported information. A study, which did not directly focus on the immune system activity, nonetheless showed support for the compensatory relationship via finding a positive association between disgust sensitivity and levels of steroids with immunosuppressive effects during pregnancy [14]. However, while this study had the advantage of using biomarkers, they were still only indirectly related to immune system activity. Finally, a smaller body of studies tested this relationship using immune markers. Most of these studies, contrary to the previously mentioned body of studies, focused on the approach of observing the so called “preparatory immune response”, with the idea that a disgust reaction, i.e. the activation of BIS, causes an upregulation of the PIS. Among the immune markers most commonly used in these studies were IL-6, a cytokine produced by white blood cells and playing a key role in the inflammatory response; secretory IgA (S-IgA), a key component of mucosal immunity, serving as a non-inflammatory immunological defence [15] or TNF-α, a potent pro-inflammatory cytokine produced during a microbial attack. In line with the proposed idea of a preparatory immune response, [16] showed an increase in levels of IL-6 after seeing photographs of infected people and [17] observed an increase in participants’ levels of salivary TNF-α and albumin in response to disgusting stimuli. The findings regarding TNF-α and albumin were then replicated and supported in a later study [18], which additionally showed an increase in body temperature in response to disgusting stimuli. Furthermore, when separating disgust- and disease-related cues (by using a set of images that primarily evoked disgust and not disease concerns and vice-versa), a significant increase in levels of salivary TNF-α for both types of stimuli was observed in people who were generally more disgust sensitive [19]. A recent study using three short videos representing different disease- and disgust-related stimuli (and one control) also showed a significant increase in S-IgA for all of the three salient stimuli [20]. Interestingly, the authors also found that the levels of S-IgA increased less (the immune reaction was not as strong) in people who had higher contamination disgust, which could also support the existence of a compensatory relationship between BIS and PIS; i.e. the people whose immune system is less active in response to disgust or disease-relevant stimuli, could have elevated contamination disgust as a compensation [20]. A different approach to testing the relationship between PIS and BIS, without trying to induce a response, was taken by Kaňková et al. [21], who assessed the activity of these systems separately. In a sample of women in their first trimester of pregnancy, the authors showed that lower levels of certain cytokines (FGF basic, Eotaxin, IFN-γ, IL-1β, IL-2, IL-4, IL-7, IL-17A, G-CSF, MCP-1, MIP-1α, PDGF-BB, IP-10, RANTES, and TNF-α) were associated with elevated disgust sensitivity, suggesting that when the immune system was not active enough, BIS activity was elevated to compensate for that [21]. However, it needs to be noted that pregnancy is an immune specific period and the observed relationship may differ in non-pregnant population. The relationship between the PIS and BIS is very complicated, with many different aspects. In our study, we aimed to add to the body of research regarding the relationship between these systems. Following the approach of Kaňková et al. [21], we did not induce disgust to observe the physiological reaction, but instead examined the relationship in its existing state. Our goal was to confirm their findings in a non-pregnant population and thus assess whether their results can be generalized regardless of reproductive status or whether they involve specific mechanisms linked to pregnancy. For the associations that were already previously observed to be significant, we took a one-sided approach to hypothesis testing. Based on this, we expected to observe evidence for a compensatory mechanism, where a less active PIS (measured by cytokine levels) would be associated with a more active BIS (measured by disgust sensitivity and general perception and behaviour related to infection and disease). Moreover, to address the limitations of text questionnaires we decided to also use tactile stimuli to evoke and measure disgust sensitivity. Methods Data collection procedure The participants were recruited as a part of a series of data collection for a larger project focused on the effect of internal and external factors on individual’s performative, physiological, and psychological characteristics, involving blood sampling (see 2.3. Blood sampling and serum analysis). They were asked whether they wanted to take part in an additional project focused on emotion and sensory perception [22]. They were then given printed study information and an informed consent to sign, including consent with using their blood serum for further analysis. The additional testing took part in a separate room or section, with the main researcher being present and giving out instructions in all data collection points. The testing involved multiple activities, including a tactile experimental task using the Tactile Disgust Stimuli Set (TDSS) [22] and filling out a set of questionnaires, asking about basic sociodemographic and overall health questions, about the participants’ disgust sensitivity and perceived vulnerability to disease (for more details, see 2.4. Measures). The data was collected in three separate time points throughout one year – in April 2022 in Olomouc, Czech Republic (S1), and in June and October 2022 in Prague, Czech Republic (S2 and S3 respectively). This research project has been approved by the Institutional Review Board of the Faculty of Science, Charles University (Approval no. 2020/27) and conducted in accordance with the Declaration of Helsinki. Sample characteristic We wanted to confirm the results of the study by Kaňková et al. [21], performed in a sample of pregnant women, on a population of non-pregnant women. Therefore, we have kept the same sample size (the sample size was determined by the size of the cytokine analysis kit) and similar inclusion criteria. In total, 78 healthy (without any serious chronic or autoimmune disorders) adult women were recruited for this study. Other inclusion criteria for recruitment were that the women were not pregnant, fluent in Czech, they filled out all or most of the questionnaires (primarily those focused on disgust sensitivity), underwent a successful blood sampling and were younger than 45. The sample characteristics are shown in Table 1. Table 1. Sample characteristics for the whole sample, as well as for the three data collection times n 78 19 30 29 Age (years; mean ± SD) range 24.4 ± 5.61 19-42 25.6±4.73 19-38 26.9±6.69 19-42 21.1±2.64 19-34 Education High school 45 (59.2%) 3 15 27 Post-secondary 3 (3.9%) 1 1 1 University 28 (36.8%) 15 12 1 missing 2 0 2 0 Place of residence 500,000 29 (38.7%) 2 17 10 missing 3 2 1 0 not-yet-known not-yet-known not-yet-known unknown Blood sampling and serum analysis The blood sampling was performed by a certified healthcare professional, with experience drawing blood. Following blood collection, the methodology used was the same as in the study by Kaňková et al. [21]. The blood sample was processed within 72 hours of collection. Serum was obtained from the blood sample and stored at -20 °C until analysed. The laboratory analyses of the serum samples were carried out at the Institute of Endocrinology in Prague in April 2023. The blood serum samples were analysed for cytokine concentrations using multiplex bead-based suspension array system (xMAP technology, Luminex Corp.) with the Bio-Plex Pro Human Cytokine 17-plex Assay (Cat. No: M5000031YV, Bio-Rad) under standard protocol. The cytokines detected with this assay are: G-CSF, GM-CSF, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p70), IL-13, IL-17A, MCP-1 (MCAF), MIP-1, TNF-α. All samples were run on a single plate. Data were evaluated by 5-parameter logistic regression with Bio-Plex Manager v. 6.1.1 (Bio-Rad). Measures Disgust Scale-Revised The Disgust Scale-Revised (DS-R) [23] is a self-report questionnaire measuring a person’s disgust sensitivity. It contains 25 items, in the form of statements, that the participants rate on a scale from 0 to 4, either depending on how much they agree with them (13 items) or how disgusting they find them (12 items). Higher score indicates higher disgust sensitivity. The questionnaire items are divided into three subscales: the core subscale (with 12 items concerned with disgust elicited by food, animals or bodily fluids), the animal reminder subscale (including 8 items related to injuries, mutilations, blood and death) and contamination subscale (with 5 items focused on interpersonal transmission of pathogens). This study used the Czech version of this questionnaire, by Polák et al. [24]. In case an answer to an item was missing from the questionnaire, the mean score of the corresponding subscale was substituted in its place. This was only done if less than one-fifth of the responses were missing from that subscale. The internal reliability for this questionnaire and its subscales, indicated by the Cronbach’s α value is 0.819 for the overall score, 0.660, 0.811 and 0.416 for the core, animal reminder and contamination subscales respectively. Three Domains of Disgust Scale The Three Domains of Disgust Scale (TDDS) [25] is another self-report tool used to measure individual disgust sensitivity. It consists of 21 items – statements, which participants rate on a scale from 0 to 6 depending on how disgusting they find them. Higher score indicates higher disgust sensitivity. The items in this questionnaire are divided into three domains: the pathogen, sexual and moral domain, each containing 7 items. In case an answer to an item was missing from this questionnaire, the mean score of its domain was substituted in its place. This was only done if less than one-fifth of the responses were missing for that particular domain. The internal reliability for this questionnaire and the three domains, indicated by the Cronbach’s α value is 0.779 for the overall questionnaire, 0.608, 0.797 and 0.873 for the pathogen, sexual and moral domains respectively. Perceived Vulnerability of Disease The Perceived Vulnerability to Disease questionnaire (PVD) [26] is used to measure an individual’s perception of their own susceptibility to illness and their protective behaviours. Participants rate, on a scale from 0 to 6, how much they agree with 15 given statements. Out of these, 7 items belong to the Perceived Infectability subscale, and 8 items belong to the Germ Aversion subscale. As before, if a response to an item was missing, the mean score of its corresponding subscale was substituted. This was done only if one-fifth or less of the responses for that subscale were missing. The internal reliability for this questionnaire and its subscales, indicated by the Cronbach’s α value is 0.743 for the overall score, 0.893 and 0.508 for the Perceived Infectability and Germ Aversion subscale respectively. Tactile Disgust Stimuli Set The TDSS [22] consists of 6 objects, meant to evoke disgust, hidden in boxes. These objects can be considered disgusting based on their tactile characteristics, while also being potentially reminiscent of specific disgusting objects or substances. The objects in this set are: soaked tapioca (fish insides), molasses on baking paper with dried fruits (flypaper with insects), bathroom tile with shaving cream (dirty bathroom), oatmeal with pear pieces (vomit), wet Play-Doh in excrement shape (excrement) and mayonnaise (pus). Participants are asked to put their hand inside, touch the object and answer a series of questions, including: “How disgusting did you find touching this object?”, “How disgusting would it be to put this object in your mouth?”, and “If you ate/licked this object, what is the probability that you would get ill?”. Responses are given on a scale from 1 (not at all/none) to 7 (very/high). Based on these responses, three scores are calculated as the sum of responses to each question from all objects: the Tactile Disgust score, Oral Disgust score, and Sickness Probability score respectively. Statistical Analyses The data was analysed using the jamovi software [27]. Firstly, using a Pearson correlation analysis controlled for age, the obtained scores of the questionnaires were correlated with each other to show the relationships between the different measures. To determine the associations between disgust and cytokine levels, we used partial Kendall correlation with age as a covariate, as the cytokine data were not normally distributed. We used the measured observed concentrations for each cytokine. In case more than half of the measurements were out of range (OOR), that particular cytokine was excluded from analysis. Overall, three cytokines were excluded from further analysis (GM-CSF, IFN-γ, IL-4). When the observed concentration for a cytokine was OOR in less than half of the measurements, a very low number was substituted in its place to allow data analysis. Nine cytokines contained substituted low values. Overall, 14 cytokines were included in further analyses. Descriptives of cytokine concentrations are included in the Supplementary material (Table S1). For those cytokines that previously showed a significant association with disgust (measured by the DS-R) in the study by Kaňková et al. [21], that analysis (for the DS-R) was performed with one-sided tests. Lastly, two Kendall correlation analyses were performed between the cytokine levels and the tactile experimental variables and the PVD questionnaire, also controlled for age. Additionally, the statistically significant results were tested for the false discovery rate, using the Benjamini-Hochberg Method, with FDR = 0.1. The data are available at: https://doi.org/10.6084/m9.figshare.29796332.v1 . Results A Pearson correlation analysis revealed moderate to strong positive intercorrelations between DS-R subscales and the pathogen domain of TDDS. The core and contamination subscales of DS-R (as well as the overall DS-R score) and the pathogen domain of TDDS were also significantly positively correlated with the Germ Aversion subscale of PVD. The sexual and moral domains of TDDS, nor the Perceived Infectability subscale of PVD, did not significantly correlate with any other measures. For more details, see Table 2. Table 2. Pearson correlation analysis between all questionnaire measures not-yet-known not-yet-known not-yet-known unknown Overall Core Animal Reminder Contamination Pathogen Sexual Moral Perceived Infectability DS-R Core 0.859*** - - - - - - - Animal reminder 0.823*** 0.487*** - - - - - - Contamination 0.627*** 0.433*** 0.328** - - - - - TDDS Pathogen 0.674*** 0.638*** 0.529*** 0.352*** - - - - Sexual 0.232* 0.185 0.186 0.179 0.270* - - - Moral -0.043 -0.040 -0.035 -0.016 -0.006 0.160 - - PVD Perceived Infectability 0.042 0.070 -0.019 0.079 0.032 -0.146 -0.084 - Germ Aversion 0.303** 0.234* 0.142 0.484*** 0.338** 0.134 0.076 0.155 Pearson’s r values are reported in the table, with significance marked as follows: * <0.05, ** <0.01 and *** <0.001. Analyses are controlled for age. For the main analysis, the possible associations between disgust measures and cytokine levels were analysed using partial Kendall correlation. For those cytokines that have shown to be significantly associated with disgust in the study by Kaňková et al. [21], specifically IL-1β, IL-2, IL-17, MCP-1, MIP-1β and TNF-α, we used one sided tests in the analysis with DS-R, with the previously observed correlation direction (negative). The results showed a significant negative correlation between cytokines IL-1β, MCP-1 and TNF-α with either core or contamination subscale of the DS-R (and in some cases also the overall DS-R score), which did not, for the most part, remain significant after a Benjamini-Hochberg correction. There was also a significant negative correlation of MIP-1β with the moral domain of TDDS, which did not withstand the correction either. Lastly, we observed significant positive associations of IL-5 and IL-12 with core and overall DS-R score, which remained mostly significant after the Benjamini-Hochberg correction. The complete results are displayed in Table 3. The results which remained significant after the additional false discovery rate tests are marked with an asterisk. The correlation analysis between the PVD questionnaire and cytokine levels showed no significant associations and neither did the correlation analysis between the tactile experimental variables and cytokine levels. The results are displayed in Table 4 and Table 5, respectively. Table 3. Kendall correlation analysis between disgust measures and cytokine levels not-yet-known not-yet-known not-yet-known unknown Core Animal Reminder Contamination Overall Pathogen Sexual Moral N = 77 N = 78 N = 78 N = 77 N = 78 N = 78 N = 77 G-CSF tau b 0.019 0.002 -0.014 0.011 0.014 -0.037 0.065 p 1.000 1.000 0.429 1.000 0.870 0.656 0.437 IL-1β tau b -0.031 -0.020 -0.164 -0.042 0.027 0.010 -0.084 p 0.345 0.400 0.017 0.294 0.750 0.822 0.314 IL-2 tau b 0.050 -0.022 -0.018 0.016 0.019 0.027 -0.000 p 1.000 0.388 0.406 1.000 0.825 0.750 0.996 IL-5 tau b 0.266 0.095 0.130 0.217 0.150 0.103 -0.049 p <0.001* 0.224 0.094 0.006* 0.067 0.208 0.546 IL-6 tau b 0.090 -0.029 -0,052 0.023 0.043 -0.056 -0.121 p 0.250 0.706 0.504 0.770 0.596 0.488 0.133 IL-7 tau b 0.050 -0.023 -0.063 0.015 0.011 -0.021 -0.154 p 1.000 0.385 0.209 1.000 0.901 0.804 0.068 IL-8 tau b 0.028 0.033 -0.035 0.029 0.075 0.014 -0.091 p 0.716 0.668 0.655 0.707 0.344 0.856 0.245 IL-10 tau b 0.049 -0.019 0.054 0.022 -0.055 0.103 -0.073 p 0.531 0.807 0.487 0.779 0.530 0.235 0.405 IL-12 (p70) tau b 0.193 0.066 0.071 0.167 0.045 -0.035 0.010 p 0.013* 0.398 0.361 0.033 0.589 0.673 0.907 IL-13 tau b 0.038 0.122 0.035 0.086 -0.047 0.103 -0.024 p 0.629 0.117 0.654 0.272 0.579 0.222 0.774 IL-17 tau b 0.058 -0.001 0.001 0.048 -0.031 0.018 -0.132 p 1.000 0.494 1.000 1.000 0.715 0.834 0.119 MCP-1 (MCAF) tau b -0.159 -0.067 -0.057 -0.139 -0.081 -0.082 0.001 p 0.021* 0.195 0.232 0.038 0.304 0.296 0.989 MIP-1β tau b 0.082 0.047 0.005 0.094 0.155 0.034 -0.165 p 1.000 1.000 1.000 1.000 0.049 0.669 0.036 TNF-α tau b 0.015 -0.050 -0.164 -0.063 -0.019 0.080 -0.089 p 1.000 0.259 0.017 0.210 0.814 0.321 0.276 not-yet-known not-yet-known not-yet-known unknown *remained significant after Benjamini-Hochberg correction. Cytokines for which one-sided tests were used are underlined. Table 4. Kendall correlation analysis between PVD and cytokine levels Overall Score Perceived Infectability Germ Aversion N = 77 N = 78 N = 77 G-CSF tau b -0.101 -0.138 -0.028 p 0.196 0.076 0.721 IL-1β tau b -0.108 -0.013 -0.141 p 0.167 0.868 0.072 IL-2 tau b -0.022 -0.073 0.028 p 0.777 0.349 0.724 IL-5 tau b 0.113 0.027 0.134 p 0.150 0.724 0.086 IL-6 tau b 0.043 0.066 0.008 p 0.582 0.396 0.920 IL-7 tau b -0.011 0.002 -0.037 p 0.887 0.977 0.632 IL-8 tau b -0.116 -0.043 -0.103 p 0.137 0.578 0.188 IL-10 tau b -0.022 -0.017 -0.015 p 0.776 0.832 0.849 IL-12 (p70) tau b 0.065 0.008 0.084 p 0.410 0.916 0.283 IL-13 tau b -0.089 -0.097 -0.076 p 0.256 0.211 0.332 IL-17 tau b -0.090 -0.140 0.004 p 0.252 0.071 0.954 MCP-1 (MCAF) tau b 0.051 0.047 -0.013 p 0.514 0.545 0.868 MIP-1β tau b -0.064 -0.034 -0.027 p 0.415 0.663 0.732 TNF-α tau b -0.076 -0.057 -0.046 p 0.332 0.462 0.554 Table 5. Kendall correlation analysis between tactile experimental variables and cytokine levels Tactile disgust Oral disgust Sickness probability N = 78 N = 78 N = 78 G-CSF tau b 0.059 -0.089 0.027 p 0.445 0.248 0.729 IL-1β tau b -0.051 -0.074 -0.124 p 0.512 0.336 0.107 IL-2 tau b 0.080 0.001 -0.011 p 0.298 0.984 0.892 IL-5 tau b 0.032 0.040 0.035 p 0.680 0.601 0.653 IL-6 tau b 0.032 -0.037 -0.084 p 0.679 0.634 0.274 IL-7 tau b 0.046 -0.043 -0.001 p 0.550 0.573 0.988 IL-8 tau b 0.040 0.066 -0.031 p 0.603 0.393 0.685 IL-10 tau b 0.016 -0.016 -0.037 p 0.840 0.836 0.629 IL-12 (p70) tau b -0.088 0.022 0.037 p 0.255 0.776 0.628 IL-13 tau b 0.047 0.029 0.005 p 0.542 0.705 0.945 IL-17 tau b -0.076 -0.057 -0.150 p 0.324 0.457 0.051 MCP-1 (MCAF) tau b -0.034 -0.022 -0.112 p 0.659 0.771 0.146 MIP-1β tau b -0.016 0.002 -0.040 p 0.839 0.979 0.603 TNF-α tau b -0.060 -0.016 -0.038 p 0.441 0.840 0.623 not-yet-known not-yet-known not-yet-known unknown Discussion In our study, we aimed to investigate the relationship between the PIS, represented here by cytokine levels and the BIS, represented by text-based disgust questionnaire scores. Based on previous research and especially a study on pregnant women using a similar design [21], we expected to observe a negative relationship between disgust and selected cytokine levels. The results of our study, however, were twofold. On one hand, in line with Kaňková et al. [21], we found the expected negative relationship between disgust sensitivity and MCP-1, IL-1β and TNF-α. On the other hand, we then also observed a positive relationship between pathogen-related disgust sensitivity and levels of MIP-1β, as well as IL-5 and IL-12, which were not measured in the previous study. We found no significant correlation between cytokine levels and PVD, nor the tactile experimental variables. We observed results that support the existence of a compensatory mechanism between BIS and PIS – if the PIS is not active or activated enough, an increase in the activity of BIS will compensate for that in order to protect the individual. In our data, we found weak negative associations between levels of pro-inflammatory TNF-α and contamination disgust, MCP-1, cytokine playing a key role in the initiation of the immune response, and core disgust, and pro-inflammatory IL-1β and contamination disgust. These findings are in line with the study on pregnant women [21] with the exception that in pregnant women, the negative association with TNF-α was observed only for core disgust. A similar pattern was also reported by Keller et al. [20], who observed a negative relationship between contamination disgust and change in S-IgA levels – people whose immune reaction, marked by change in S-IgA levels, was not as strong had higher contamination disgust. This evidence is also in line with our results regarding TNF-α and suggests that a similar compensatory mechanism could be occurring in cases of both immune markers. However, at the same time, in a different study [19], which used TNF-α as an immune marker, showed an increase in TNF-α levels in response to disgust- and disease- cues in more disgust sensitive people, suggesting evidence for a simultaneous activation of both BIS and PIS. It can only be noted that Keller et al. [20] also found these results using the DS-R, a questionnaire for which we observed our significant associations, while Stevenson et al. [19] observed their results for the TDDS, which did not show any significant negative association with the TNF-α, MCP-1 or IL-1β in our study. Lastly, it is necessary to say, that even though the results of our observed negative association were significant in a classic partial Kendall correlation, they did not withstand the Benjamini-Hochberg correction, and the contamination subscale of DS-R showed low reliability. Therefore, any possible interpretations should therefore be approached with caution. While some of the results align with those reported by Kaňková et al. [21], several cytokines, namely IFN-γ, IL-2, IL-4, IL-7 and IL-17, showed a significant negative association with disgust in their sample of pregnant women, but not in our sample of non-pregnant women. This discrepancy highlights the importance of considering the unique immunological context of pregnancy. During pregnancy, the maternal immune system undergoes numerous changes and modifications [28,29], cytokine levels can be present in different concentrations and the dynamic between PIS and BIS can change during this time. Indeed a study has shown that levels of IFN-γ and IL-2 differed significantly between non-pregnant and pregnant women [30]. A different longitudinal study showed dynamic changes in cytokine levels throughout the whole pregnancy, with levels of proinflammatory cytokines being highest in first and third trimester [31]. Levels of many cytokines (such as IL-2 or IL-17) seem to increase from the first to third trimester [32]. This suggests that the compensatory mechanisms between PIS and BIS may become more pronounced during pregnancy, especially in the first trimester, serving to maintain protection against pathogens that could compromise foetal development. We have also observed a positive correlation between disgust and cytokine levels, specifically between core disgust, overall DS-R and IL-5; core disgust, overall DS-R and IL-12; and the pathogen disgust domain of TDDS and MIP-1β. The results regarding IL-5 and the association between core disgust and IL-12 also remained significant even after the Benjamini-Hochberg correction. IL-5 is expressed in eosinophils, which play a key role in the pulmonary inflammation associated with allergies [33,34]. IL-12 has also been shown to be higher in patients with asthma [35] and its role in allergic airway inflammation during the effector phase on an animal model [36]. While also still being proinflammatory cytokines, IL-5 and IL-12 have also been shown to be associated with neuroticism and with the severity of depressive symptoms [37]. Patients with diagnosed major depressive disorders had higher levels of IL-12 than healthy controls [38]. In a study on pregnant women, IL-12 has been found to be associated with prenatal depressive and overall anxiety symptoms and IL-5 with depressive symptoms [39]. There is a connection between these two areas of research – i.e., between the role of IL-5 and IL-12 in mental health and in respiratory problems – studies have shown a significant positive association between the prevalence of anxiety and asthma [40,41]. In order to understand how these relationships are all interconnected, we need to focus on the last variable – disgust. Disgust, while a part of the BIS, is also often positively associated with mental health. Specifically, it has been shown to be associated with fears/phobias [42], depression [43], obsessive-compulsive disorder [44] and other anxiety disorders [45,46]. Elevated disgust in the first trimester has also been shown to predict higher anxiety in the third trimester [47]. Personality-wise, it has also been shown to be associated with neuroticism [48]. The observed positive association between disgust and cytokines could therefore be related to the changes in mental health and even tied to respiratory issues. Interestingly, we found no significant association between cytokine levels and pathogen avoidance motivation measured by the PVD. These findings are consistent with some of the previous studies that also observed primarily associations between disgust and immune markers, but did not find significant links with PVD [19,20]. On the other hand, a study by Gassen et al. [49], which focused specially on pathogen avoidance motivation (measured by the Germ Aversion subscale of PVD), showed that individuals with higher pathogen avoidance motivation had lower levels of plasma IL-6 measured in vivo and less spontaneous release of IL-1β, IL-6, and TNF-α in vitro. However, this higher pathogen avoidance did not predict a decreased release in response to immune stimulation, suggesting that increase BIS activity does not necessarily have to predict lower PIS activity. Moreover, the study showed that people with higher pathogen avoidance motivation also had lower oxidative stress. Their discussion of these results brings another possible explanation to the mechanisms between BIS and PIS – that higher BIS activity could protect the body from the harmful effects of inflammation and oxidative stress, while the activity of PIS in times of infection would remain normal. Although our results did not show any significant negative relation between pathogen avoidance and cytokine levels, which would be in line with this explanation, we did observe this relationship between disgust and levels of IL-1β, TNF-α and MCP-1. This could suggest that while pathogen avoidance and disgust are both different components of the BIS, they may represent different mechanisms of behavioural immunity. It is worth noting that the study by Gassen et al. [49] included a much younger (student) sample and over half of the participants were men. It is possible that different behavioural strategies are differently represented in men and women and in different age groups. We also didn’t find any associations between the variables related to tactile disgust and cytokine levels. In the original study establishing the TDSS tactile measure [22], which had an overlapping sample with the current study, the strongest association was with the animal reminder subscale of DS-R. This subscale also turned out to play a significant role in how the different tactile objects were rated (along with the Germ Aversion subscale of PVD). The authors discuss this via the connection between body vulnerability and body envelope concerns (represented by the animal reminder disgust) and the proximate modality of touch. In this study, no cytokine levels were associated with animal reminder disgust. Overall, it seems that tactile disgust is, similarly to the pathogen avoidance motivation mentioned above, connected to a different component of the BIS. Therefore, tactile disgust measures might not be the most suitable for exploring the relationship between BIS and PIS. Measures using other sensory modalities, such as smell, which is more involved in pathogen detection from the environment than touch, could be a better alternative to use in future studies. Regarding to other domains of disgust, our results did not show any statistically significant association between cytokine levels and sexual disgust. While sexual disgust does partially serve a pathogen-protective role (mainly against sexually transmitted diseases) its main function is to protect from sexual behaviour that could lead to lower reproductive success [25]. Therefore, we did not expect to see a direct correlation with immune markers. On the other hand, we observed a significant negative association of MIP-1β with the moral disgust domain of TDDS. While no studies directly focused on the link between moral disgust and PIS activity, a study [50] showed that both pathogen and moral disgust was associated with ratings of both pathogen-related and moral-related mitigation actions in the context of the Covid-19 pandemic (people who had higher disgust were more likely to rate norm violations as both morally wrong and more likely to lead to Covid infection), showing that there is some possible overlap in these two types of disgust in regard to BIS. It is important to note that, in contrast to negative association between MIP-1β and moral disgust, we also found an opposite (positive) association between MIP-1β and the pathogen disgust domain, which does not align with the aforementioned interpretation. On the other hand, all results pertaining to TDDS were weak and did not withstand the Benjamini-Hochberg correction. Strengths and limitations One of the strengths of this study is the simultaneous use of multiple disgust questionnaires, as well as the use of a tactile experimental task to evoke and measure disgust. This allows us to better understand how different components of BIS relate to markers of immunity. The use of a laboratory kit for the analysis of 17 cytokines allowed us to examine not only the expected associations, but also new relationships, taking into account different aspects of disgust. One limitation of the study may be the relatively healthy sample of women without serious medical conditions, which could have reduced variability in immune functioning and may have contributed to the lack of certain associations that might be detectable in a more immunologically diverse or immunosuppressed population. This interpretation is further supported by our findings, where the strongest correlations were observed with cytokines that are likely more closely related to mental health status than to specific immune responses. Conclusion This study adds to the body of research regarding the relationship between the physiological immune system and the behavioural immune system. Similarly to a previous study in pregnant women, our results provide weak evidence for the compensatory-type relationship, as some cytokine levels were negatively associated with disgust sensitivity. Although our findings allow for some generalization from pregnant to non-pregnant populations, several cytokines that showed significant negative associations with disgust in pregnant women did not show the same pattern in our sample of non-pregnant women, further suggesting the unique immunological context of pregnancy. Moreover, we found strong positive associations between disgust sensitivity and cytokine levels that have repeatedly been shown to be associated with respiratory conditions, such as asthma or allergies, as well as to mental health, such as neuroticism or anxiety. The positive association observed in our study may therefore reflect this interconnected relationship between disgust, mental health and respiratory functioning. Lastly, we investigated whether cytokine levels were associated with other aspects of the behavioural immune system, specifically the vulnerability to disease and pathogen avoidance motivation, as well as disgust in response to tactile stimuli. None of these measures showed any significant association with the observed cytokine levels. This highlights the complexity and heterogeneity of the mechanisms that constitute the behavioural immune system and disgust sensitivity itself, suggesting that different components of behavioural protection may be differently linked to physiological processes. Funding This work was supported by the Czech Science Foundation, project GA CR (23-05519S). Conflict of interest The authors declare no conflict of interest. not-yet-known not-yet-known not-yet-known unknown Data availability The open data are available at: https://doi.org/10.6084/m9.figshare.29796332.v1 Acknowledgements We would like to thank the participants for their time and effort in taking part of this study. We would also like to thank Martin Hůla for his help during data collection. References [1] Schaller M, Duncan LA. The behavioral immune system: Its evolution and social psychological implications. Evol. Soc. Mind Evol. Psychol. Soc. 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Authors Affiliations Daniela Dlouhá 0000-0002-2968-1992 [email protected] Univerzita Karlova Biologicka sekce View all articles by this author Josef Včelák Endokrinologicky ustav View all articles by this author Šárka Kaňková Univerzita Karlova Biologicka sekce View all articles by this author Metrics & Citations Metrics Article Usage 244 views 92 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation Daniela Dlouhá, Josef Včelák, Šárka Kaňková. The relationship between disgust sensitivity and serum cytokine levels in non-pregnant women: a generalization of previous results on pregnant population. Authorea . 19 August 2025. DOI: https://doi.org/10.22541/au.175558739.94517887/v1 If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. Simply select your manager software from the list below and click Download. 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