Heterologous immune responses in health and disease

Immunological memory and tolerance represent major achievements and advantages of adaptive immunity. Organisms bearing adaptive immunity display prominent competitive advantages in the fight against infections. Memory immune cells are preserved for decades and are able to repel a second attack of an infectious agent. However, studies performed in the XXI century have shown that even unrelated pathogens may be quickly and effectively destroyed by memory cells. This type of response is called heterologous so that heterologous immune response is mainly typical to viral infections and other intracellular infections, where T-cells play a lead role in protection. This review will discuss various mechanisms involved in implementing T-cell cross-reactivity, describe molecular prerequisites for heterologous T-cell responses. Experimental evidence of memory T-cell potential to heterologous immune response in mouse models and in human infections are also discussed. Heterologous immune response is an important immune arm in adults and the elderly when the yield of naive cells to the periphery declines due to thymus involution. Along with obvious advantages, heterologous immune response leads to imbalanced memory T-cell repertoire, replacement of immunodominant epitopes with minor ones allowing viruses to evade immune response that results in virus persistence, or, conversely, fulminant infection course. Another threat of heterologous immune response due to switch in dominant repertoire of recognizable epitopes is presented by random self-epitope recognition, which can lead to development of autoimmune pathology. Heterologous immunity can also disrupt drug-induced tolerance in organ and tissue transplants and lead to graft rejection. Heterologous immune response should be taken into consideration while developing and using new vaccines, especially in adults and the elderly

Interleukin-36 family as a novel regulator of inflammation in the barrier tissues

The interleukin-36 (IL-36) family was discerned in the superfamily of interleukin-1 (IL-1) ten years ago. This family includes three isoforms of IL-36α, IL-36β, IL-36γ, which have pro-inflammatory activity and a specific receptor antagonist, IL-36ra, which implements anti-inflammatory function. All of them bind to the same IL-1R6 receptor. The pro-inflammatory isoforms also involve an accessory IL-1RAcP protein into signaling; resulting into conduction of a signal into the cell via the assembling heterodimer receptor. In contrast, IL-36ra inhibits the formation of a heterodimer and blocks the signal transmission. The cytokines of the IL-36 family and appropriate receptors are normally expressed on epithelial cells in barrier tissues such as the respiratory, intestinal tract and skin. Like all cytokines of the IL-1 superfamily, IL-36 is synthesized as inactive form and requires activation, but not due to caspases, but being mediated by neutrophil enzymes, such as cathepsin G, proteinase-3, and elastase, which are constantly present in barrier tissues. In this regard, IL-36 is involved in homeostasis of barrier tissues. Apparently, the IL-36 cytokine system appeared in response to the developing ability of some microorganisms to avoid immune recognition and activation of innate immune response, and, in particular, the IL-1 pro-inflammatory system. An imbalance between the pro- and anti-inflammatory pathways readily causes inflammation in the corresponding tissue. This review discusses participation of cytokines from the IL-36 family in homeostasis of barrier tissues, as well as potential role of the IL-36 family in pathogenesis of bacterial, viral, and fungal skin diseases, atopic dermatitis, autoimmune diseases, such as rheumatoid arthritis, systemic lupus erythematosus, Sjogren's syndrome, ulcerative colitis and Crohn's disease. The role of IL-36 family cytokines in the immunopathogenesis of psoriasis has been well studied. This review is presenting the modern ideas about immune pathogenesis of psoriasis. The special role of cytokines from the IL-36 family was shown both for induction of psoriatic inflammation and evolving a positive feedback loop that supports and enhances the immune component of inflammation, which leads to progression of the disease. Moreover, modern methods of treating psoriasis are discussed, in particular, a possible promising approach to IL-36 blockade, or usage of recombinant IL-36ra for the treatment of psoriatic patients. Experimental studies in this area in mice provide some grounds for optimism

Gating strategy for plasmablast enumeration after hepatitis B vaccination

B cell stimulation develops upon vaccination, thus causing occurrence of activated B cells (plasmoblasts) in bloodstream. Similar cells are also observed in some viral infections. The contents of plasmablasts may be a marker of successful vaccination, or a diagnostic feature of ongoing infection. The plasmablasts are normally represented by a small cell subpopulation which is not easy to detect. A study was performed with 15 healthy volunteers who were subjected to a single immunization with a recombinant vaccine against hepatitis B virus. To identify the plasmablasts, we have used labeled antibodies prepared in our laboratory. These reagents were previously validated for counting the plasmablasts. Different gating strategies for plasmablast gating have been compared. Upon staining of lymphocytes from immunized volunteers, we observed a distinct cluster of plasmablasts with CD27++CD38++ phenotype using the following antibody set: CD19-PE, CD3/CD14/CD16-FITC, CD27-PC5.5 and CD38-PC7. Inclusion of a CD20-FITC antibody into the panel caused an increase of CD27++CD38++ plasmablast ratio among CD19+ lymphocytes to > 60%. Upon substitution of CD38 antibody by anti-CD71, a distinct plasmablast cluster was again revealed, which contained ca. 5 per cent В cells. Two strategies for the plasmablast gating using the CD27/ CD38 and CD27/CD71 combinations were compared in dynamics with lymphocyte samples from a single vaccinated volunteer. When applying the CD27/CD38 combination, a sharp and pronounced plasmablast peak was registered on day 7 post-vaccination. With CD27/CD71 combination, the peak was extended between day 7 and day 14 following immunization. Hence, time kinetics of the CD27+CD71+ population proved to be different from occurrence of classic plasmablasts with CD27++CD38++ phenotype. This finding suggests that the CD27++CD71+population contains both plasmablasts and other types of activated B cells. A minor HBV surface antigen was prepared and labeled with phycoerythrin (HBsAg-PE), thus allowing to quantify the antigen-specific plasmablasts. The results of HBsAg-PE-based detection of antigen-specific cells were in compliance with the data obtained by ELISpot technique. At the present time, we use the original plasmablast gating technique for detection of activated B cells in SARS-CoV-2 infection. At the next step, this technique will be applied to sorting of antigen-specific B cells, thus permitting sequencing of Ig genes and design of novel human antibodies against viral antigens.B cell stimulation develops upon vaccination, thus causing occurrence of activated B cells (plasmoblasts) in bloodstream. Similar cells are also observed in some viral infections. The contents of plasmablasts may be a marker of successful vaccination, or a diagnostic feature of ongoing infection. The plasmablasts are normally represented by a small cell subpopulation which is not easy to detect. A study was performed with 15 healthy volunteers who were subjected to a single immunization with a recombinant vaccine against hepatitis B virus. To identify the plasmablasts, we have used labeled antibodies prepared in our laboratory. These reagents were previously validated for counting the plasmablasts. Different gating strategies for plasmablast gating have been compared. Upon staining of lymphocytes from immunized volunteers, we observed a distinct cluster of plasmablasts with CD27++CD38++ phenotype using the following antibody set: CD19-PE, CD3/CD14/CD16-FITC, CD27-PC5.5 and CD38-PC7. Inclusion of a CD20-FITC antibody into the panel caused an increase of CD27++CD38++ plasmablast ratio among CD19+ lymphocytes to > 60%. Upon substitution of CD38 antibody by anti-CD71, a distinct plasmablast cluster was again revealed, which contained ca. 5 per cent В cells. Two strategies for the plasmablast gating using the CD27/ CD38 and CD27/CD71 combinations were compared in dynamics with lymphocyte samples from a single vaccinated volunteer. When applying the CD27/CD38 combination, a sharp and pronounced plasmablast peak was registered on day 7 post-vaccination. With CD27/CD71 combination, the peak was extended between day 7 and day 14 following immunization. Hence, time kinetics of the CD27+CD71+ population proved to be different from occurrence of classic plasmablasts with CD27++CD38++ phenotype. This finding suggests that the CD27++CD71+ population contains both plasmablasts and other types of activated B cells. A minor HBV surface antigen was prepared and labeled with phycoerythrin (HBsAg-PE), thus allowing to quantify the antigen-specific plasmablasts. The results of HBsAg-PE-based detection of antigen-specific cells were in compliance with the data obtained by ELISpot technique. At the present time, we use the original plasmablast gating technique for detection of activated B cells in SARS-CoV-2 infection. At the next step, this technique will be applied to sorting of antigen-specific B cells, thus permitting sequencing of Ig genes and design of novel human antibodies against viral antigens

CHANGES IN THE CAPILLARY AND VENOUS BLOOD CYTOKINE PROFILE OF PATIENTS WITH PSORIASIS DEPENDING ON THE TREATMENT

ABSTRACT. Psoriasis is a chronic autoimmune skin disease involving T-cell immunity. The interleukin (IL)-23/IL-17/IL-22 cytokine axis is key in the immunopathogenesis of psoriasis. The role of the IL-36 subfamily regulating inflammation in the skin is shown. Topical preparations are used to treat psoriasis. Objective: to study changes in the cytokine profile of venous and capillary blood taken near the focus of psoriatic inflammation, depending on the treatment with topical preparations. 40 patients with psoriasis, mean age 43.7 years, were examined. Group 1a (20 people) received local treatment with mometasone, Group 1b (20 people) received topical gel containing IL-36 receptor antagonist. 20 healthy people, mean age 46.6 years, consisted the control group 2. Capillary blood was collected from a finger, in patients near the lesion 200 μl in a microvette with EDTA. Venous blood was taken from the cubital vein 3 ml into a vacuum tube with EDTA. The concentration of 15 cytokines in blood plasma was tested by the multiplex method (MagPix, BioRad, USA). The effectiveness of therapy was assessed using the PASI and DLQI indices. At the end of treatment (day 14), the PASI and DLQI indices significantly decreased in both groups. On the 28th day, the PASI index in Group 1a returned to its original level, in group 1b it remained steadily reduced. Before treatment, the levels of all cytokines except IL-10 in the capillary blood of patients with psoriasis were significantly increased compared to Group 2, and the levels of 5 cytokines were increased in the venous blood. After 14 days in Group 1a, the levels of IL-1, IL-4, IL-6, IL-21, IL-22, IL-23, IL-25, IL-33 significantly decreased in capillary blood, and only IL-17F, IL-21, IL-33 and TNF in the venous blood. On the 28th day, the concentrations of almost all cytokines returned to their original level. In Group 1b, on the 14th day, the levels of IFN-γ, IL-1, IL-4, IL-17F, IL-21, IL-22, IL-23, IL-25, IL-33 significantly decreased in capillary blood, and in venous blood - IFN-γ, IL-21, IL-22, IL-23, IL-33. On the 28th day, the concentration continued to decrease, or the level of these cytokines remained reduced, and IL-6 significantly decreased in the vein. Thus, the method for determining the profile of capillary blood cytokines from the area of ​​psoriatic lesions can be used to monitor the effect of treatment in patients with psoriasis

Comparing humoral immune response in adult measles patients and measles vaccinated subjects

Introduction. The implementation of the WHO Measles Elimination Program has yielded serious results, but in recent years an increase in the incidence rate of this infection has been observed. In particular, according to the WHO, in 2019 vs. 2018 measles morbidity was elevated by 3-fold worldwide. While investigating measles outbreaks among patients, apart from unvaccinated subjects, a substantial group of adults vaccinated in childhood was distinguished. The aim of this work was to examine the characteristics of humoral measles immunity in adult measles patients as well as subjects after measles vaccination. Materials and methods. 50 adult measles patients aged 20 to 55 years were examined. In all patients, the diagnosis was confirmed clinically and by laboratory assays by detecting measles IgM antibodies. The second group consisted of 50 conditionally healthy seronegative age-matched adults, vaccinated with the live measles vaccine (Microgen, Russia). Peripheral blood samples were collected from the cubital vein in total volume of 4 ml on 6±1 day after the onset of rash in patients as well as 6 weeks after vaccination. Specific measles antibodies and their avidity were determined by ELISA using the commercial Avidity: Anti-Measles Viruses ELISA/IgG kit (Euroimmun, Germany). Results. It was shown that people aged 20—35 years more likely suffered from measles than elderly. And it was in this age group that healthy measles seronegative individuals were more abundant. Among vaccinees, 44% responded to vaccination with the primary type of immune response, and 56% responded with the secondary type, while among measles patients, 34% and 66% responded with the primary and secondary type, respectively, as follows from the spectrum of specific IgG subclasses and the antibody avidity assay. The secondary type of immune response indicates that these subjects were apparently vaccinated against measles in childhood, but lost with time long-lived plasma cells producing protective antibodies. While comparing the parameters of specific humoral immunity in groups with acute measles infection (day 6 from the onset of rash) and early convalescents (3 weeks after the onset of rash), it was shown that the level of specific IgG increased threefold in early convalescents (p < 0.01) compared with those at acute phase. The level of specific IgA, on the contrary, decreased from 73.44 (69—75.3) Me/ml to 48.64 (45—56.4) Me/ml, but remained very high. At the same time, the spectrum of specific IgG subclasses shifted from primary immune response (high IgG3 and low IgG1) to secondary response (low IgG3 and high IgG1), which is typical for the response of emerging memory B cells

Diagnostic value of anti-GP2 antibodies determined in serum and coprofiltrates in children with inflammatory bowel disease

Inflammatory bowel diseases (IBD), such  as Crohn’s disease (CD) and  ulcerative colitis (UC), are characterized by chronically recurring inflammation of intestinal wall and are associated with a significant decrease in the  quality  of life. A spectrum of genetic  variants  associated with  Crohn’s disease  is described. Intestinal dysbiosis (DB)  may be the triggering factor of the disease. Glycoprotein 2 (GP2), the main protein of pancreatic zymogen  granules, is secreted  into the intestines with digestive enzymes.  Anti-GP2 antibodies were found in the serum of patients with CD.  The aim of the present  study was to investigate  the levels of anti-GP2 antibodies in serum  and feces of children with IBD  compared with the DB group.  Serums  and coprofiltrates from 110 children (64 boys and 46 girls) at the age of 12.3 (2.6-17.9) years were studied; 36 patients with CD, 30 patients with UC.  A comparison group consisted of 44 patients with DB. IgG and IgA antibodies against GP2 were tested with ELISA. Nonparametric statistics methods are applied, the results are presented as percentages and medians (Me (Q0.25-Q0.75)). The serum levels of anti-GP2 IgA antibodies were 9.97 (3.35-13.45) U/ml for the CD patients, 6.08 (2.71-14.26) U/ml for UC and 2. 94 (2.29-6.41) U/ml for DB. The levels of anti-GP2 IgG antibodies in serum were 6.16 (3.26-18.4) U/ml for CD, 5.26 (2.97-7.52) U/ml for UC, and for DB 5.23 (2.53-8.85) U/ml. The cut-off  threshold concentration for anti-GP2 IgG antibodies was 13.8 U/ml, with sensitivity of 63.2%, specificity 100%, and for IgA 5.63 U/ml, with sensitivity of 60.5% and specificity of 78.8%, thus being lower than the calculated cut-off  for adults (20 U/ml). The levels of anti-GP2 IgG in coprofiltrates in children of comparison group  were 1.99 (1.26-3.04) U/ml; in the  patients with CD, 23.5 (16.15-29.3) U/ml, and  in children with UC, 20.45 (13.63-25.5) units/ml (p < 0.001). The cut-off  value amounted 8.0 U/ml, with 100% sensitivity  and  100% specificity.  Concentrations of anti-GP2 IgA in coprofiltrates of patients with IBD  did not significantly  differ from DB patients. Moreover, the concentration of sIgA in the coprofiltrates of patients with IBD  was significantly  higher than  their level in DB group. The anti-GP2 IgA/sIgA  ratio was significantly lower in patients with CD (0.326 (0.23-0.512)), and UC (0.327 (0.205-0.435)), than in patients with DB (2.332 (1.575-3.523)) (p < 0.001);  the cut-off  level was 0.784, with a sensitivity of 97.7% and specificity  of 98.6%. It is discussed, whether fecal anti-GP2 IgA antibodies should  be considered as protective, supporting intestinal homeostasis, whereas anti-GP2 IgG antibodies are pathogenetically significant  for development of IBD.  Thus, using a non-invasive method for determining anti-GP2 antibodies in stool, when exceeding the cut-off for IgG, and reduction of IgA/sIgA ratio below the cut-off, one may differentiate IBD from DB with a similar symptoms at the onset of disease, with 100% sensitivity and 100% specificity

Formation of humoral and cellular immunity to measles vaccine in adults

Despite adherence to the policy of mass measles vaccination in the majority of countries, this infection still remains far from being fully eradicated. Measles outbreaks are reported worldwide, when the vast majority of cases are recorded in subjects of 18—35 years of age. Studies on assessing measles IgG antibody level in different regions of Russia reveal increased percentage of measles seronegative subjects among young adults. Current study was aimed at investigating formation of humoral and cellular immunity after measles vaccination in seronegative adults aged 18 to 30 years old. There were enrolled 50 measles seronegative healthy volunteers aged 18 to 30 years old. Level of anti-measles IgM and IgG antibodies was measured by ELISA (Vector-Best, Russia). Subclasses of measles specific IgG antibodies were analyzed by ELISA, by replacing IgG conjugate for IgG1, IgG2, IgG3, IgG4 conjugates, whereas measles specific IgA antibodies were estimated by ELISA with IgA conjugate (Polygnost, Russia) at a concentration of 1 μg/ml. Antibody avidity was assessed by ELISA (Euroimmun, Germany). Cell-mediated measles immunity was estimated by CD107a surface expression on CD8hi T cell subset stimulated by measles virus-derived antigens. A specific cellular response to measles antigens before vaccination was detected in 50% of examined subjects, whereas 40% samples showed no signs of cellular immune response, with 10% of remaining cases described as equivocal. It was found that 6 weeks after vaccination all vaccinated subjects developed measles specific IgG antibodies at protective level reaching 1.33 (0.85—1.82) IU/ml [Me (LQ—UQ)]. Anti-measles IgA antibodies were of 0.655 (0.423—1.208) IU/ml [Me (LQ—UQ)]. However, no measles specific IgM antibodies were detected 6 weeks after vaccination. In addition, primary type of immune response (dominant low-avidity anti-measles antibodies IgG3 subclass) to measles vaccination was observed in 24 out of 50 subjects, whereas 26 subjects developed secondary type of immune response (high-avidity anti-measles antibodies dominated by IgG1 subclass). A measles specific cellular immune response was observed in 47 of the 50 examined subjects, and in 3 volunteers it was equivocal. Further analysis revealed a cohort of subjects who were not vaccinated against measles (18 subjects), although 60% of them provided medical record on previous dual measles vaccination occurred in childhood. Another cohort consisted of subjects who had medical record of measles vaccination in childhood (32 subjects), but lost protective measles antibodies produced by plasma cells (23 subjects), and memory T cells (3 subjects), or measles antibodies and memory B cells (6 subjects) over time. Such pattern evidences that measles-specific cellular and humoral arms immune responses were developed and maintained independently of each other

Evaluation of age-related distribution of measles cases with primary and secondary immune response in Russian Federation, 2010-2016

In 2010—2016, blood serum samples were examined from 5539 patients, aged < 1—60 years, with clinically and laboratory confirmed measles. Primary or secondary type of immune response was determined for all measles cases. Studies were performed with children aged < 1—14 years (2381), adolescents, 15—17 years old (189), and adults aged 18—60 years (2969). Serum measles-specific IgM antibodies were measured by “VektoKor’ IgM” ELISA test system (Russia), concentration and avidity of specific IgG — by using “Anti-Measles Viruses ELISA/IgG” and “Avidity: Anti-Measles Viruses ELISA/ IgG” (Euroimmun, Germany). Primary immune response was identified based on the presence of serum measles-specific low avidity IgM and IgG antibodies, whereas secondary immune response was characterized by detecting high avidity IgM and IgG antibodies at concentration of ≥ 5.0 IU/ml. Analyzing measles-specific IgM antibodies in 2010—2016 demonstrated that measles morbidity was mainly due to children, aged 1—2 years reaching up to 39.9% of the total number of children with measles aged < 1—14 years as well as adults aged 18—40 years old comprising as high as 80.1% total number of patients aged 15—60 years. Serum measles-specific IgG testing showed that in 15.0% of cases they were detected at concentration of ≥ 5.0 IU/ml. Further serum dilution resulted in finding IgG titer ranging within 8.5—45.0 IU/ml (21.4+0.36) and high avidity antibodies in 80—100% (92.5+0.2) cases. The remaining 85.0% cases found low avidity measles-specific IgG antibodies (< 30%) at concentration of 0.2—3.46 IU/ml (1.73+0.03). An age-related analysis of our data demonstrated that all children under 14 with laboratory-confirmed measles developed primary immune response. Moreover, in 73.7% of measles patients aged 15—60 with primary immune response measles might be prevented by timely vaccination, whereas persons with “vaccine failure” comprised 26.3%. In 2010 (0.09 per 100,000 subjects) and 2016 (0.12 per 100,000 subjects), frequency of patients with “vaccine failure” during relative epidemic well-being was 35.3% and 18.2%, respectively, exceeding 9.9% (p < 0.001) serving as a hallmark 2014 high measles incidence rate (3.24 per 100,000 subjects).The data obtained indicate that measles virus circulate among people with “vaccine failure,” which may account for potential to spread and infect unprotected population cohorts as well as cause measles outbreaks during periods of epidemic well-being

Assessment of serological tests for antibodies to different antigens of the SARS-CoV-2 virus: comparison of six immunoassays

The new coronavirus SARS-CoV-2 has become a global challenge to medicine and, in particular, laboratory diagnostics. The study of the antibodies’ level to SARS-CoV-2 can be used as a confirmation test in the diagnosis of a disease, but it becomes of paramount importance in assessing population immunity resulting from a disease or vaccination, as well as in selection of convalescent plasma donors. The kits developed in our country and abroad for detecting antibodies to the SARS-CoV-2 virus differ both in the methods of testing and in the used coronavirus antigens to which the antibodies are directed. The aim of this study was to compare the diagnostic sensitivity and specificity of five kits for the detection of IgG antibodies to the SARS-CoV-2 virus, based on different diagnostic methods. Serum samples from 137 COVID-19 convalescents and 166 donors of blood and its components were examined. The control group consisted of 50 blood sera collected at the beginning of 2019 and 19 sera collected in 2018 (before the advent of the SARS-CoV-2 virus) and stored at -70 °C. Testing was carried out in analytical systems: rapid test “COVID-19 IgM/IgG Rapid Test (Colloidal Gold)” (China), on an automatic immunochemical analyzer Abbott Architect™ i2000 and kit “SARS-CoV2-IgG” (Abbot, Chicago , IL USA), by the chemiluminescence method using an automatic analyzer of the CL series and kits of the “Mindray” company (China) “SARS-CoV-2 IgM” and “SARS-CoV-2 IgG” and by the enzyme immunoassay method on the kits of the companies “Diagnostic Systems” Ltd (Russia, Nizhny Novgorod) “DS-IFA-ANTI-SARS-CoV-2-G”, “Xema” Ltd (Federal State Budgetary Institution “National Medical Research Center of Hematology” of the Ministry of Health of Russia) “SARS-CoV-2-IgG-IFA” and “Vector-Best” CJSC (Russia, Novosibirsk)” SARS-COV-2-IgM-IFA-BEST” and “SARS-COV-2-IgG-IFABEST”. When comparing the results of testing 137 plasma samples on the Vector-Best and Mindray kits for IgG antibodies, 127 samples were positive, 7 samples were negative on both kits, the discrepancy was 2.2%. In the study of IgM antibodies, 32.1% were positive, and 52.6% were negative in both kits. The discrepancy rate was 15.3%. Out of 166 samples, 1 serum (0.6%) was negative in 5 kits. On the Mindray kit, IgG antibodies to the antigens of the SARS-CoV-2 virus were detected in 165 samples (99.4%), on Vector-Best – in 164 sera (98.8%), on Diagnostic systems – in 151 (90.96%), on Xema – in 154 (92.8%), and on Abbott – in 155 samples (93.4%). At the same time, 135 (81.33%) samples were positive in all kits, while 30 samples had discordant results (18.07%), and in 9 sera, specific IgG was not detected in 2 or more kits. ROC analysis revealed a high diagnostic value of all tested kits (AUC from 0.908 to 0.998), which indicates a high quality of the separation model of positive and negative samples (p < 0.001). With the cut-off set by the manufacturers, the sensitivity and specificity ranged from 82.8% and 93.3% for the Diagnostic Systems kit to 99.4% and 95.8% for the VectorBest kit. The calculated correlation coefficients were higher between kits with a similar composition of the antigen used in the kits; therefore, it is better to monitor the dynamics of antibodies by diagnostic kits from the same manufacturer

Mononuclear subsets and cytokine profile of venous and capillary blood in patients with psoriasis and healthy people

Psoriasis is considered an autoimmune disease with a predominantly cellular mechanism for the development of disorder. Studies in immune pathogenesis of psoriasis were performed either in animal model, which is not just similar to humans, or the data were obtained in patients by means of skin window method, which is traumatic, or by examining venous blood. However, it is difficult to discern parameters of the local immune response in venous blood samples. We have attempted to find an adequate method which would be convenient both for the patient and for the researcher, in order to assess local immune processes occurring in the skin affected by psoriasis. We examined 20 patients with a verified diagnosis of psoriasis, the average age was 44.3 years. The control group included 15 healthy adults, with average age of 46.6 years. Capillary blood was taken by fingerprick, whereas, in psoriatic patients, the samples were taken near the psoriatic lesion at a final volume of 400 μL in two microvettes. Venous blood (3 mL) was taken from the cubital vein into a vacuum tube with EDTA. Clinical analysis of venous and capillary blood was performed in automated hematological analyzer. Immunophenotyping was performed by four-color staining of whole capillary and venous blood followed by lysis of erythrocytes. Cytofluorometry was performed using techniques and reagents from BD Biosciences (USA). Plasma cytokines were determined by multiplex approach (MagPix, BioRad, USA). Upon clinical analysis of blood, the difference between capillary and venous blood was not found, either in healthy group, or among patients with psoriasis. In healthy people, the subsets of mononuclear cells, did not differ between venous and capillary blood. The samples of capillary and venous blood in the patients with psoriasis showed significantly increased levels of double-positive lymphocytes (CD45RA+/CD45R0+), B lymphocytes and NKT lymphocytes (both for relative and and absolute values). A significant increase in the percentage of naive T lymphocytes, activated helpers (Thact) and Treg, as well as B1 cells and Breg, and a significant decrease in B2 lymphocytes was registered in capillary blood of the patients with psoriasis. In venous blood samples from psoriatic patients, only a significant increase in Thact, Treg, and Breg was revealed. In the capillary blood of patients with psoriasis, we found a significant increase in the levels of non-classical M2 monocytes and inflammatory Minfl monocytes, and a decrease in classical M1 monocyte levels; in venous blood of psoriatic patients, only an increase in inflammatory Minfl monocytes was revealed. In capillary blood, all the studied cytokines in psoriasis patients significantly exceeded the levels of corresponding cytokines in healthy controls, except of IL-10. The levels of this cytokine did not differ from healthy group. In venous blood, the levels of most studied cytokines in the group of patients with psoriasis did not differ from the group of healthy ones. Approximately two-fold increase was revealed for IL-4, IL-21, IL-23 and TNF. First, the subsets of mononuclear cells and the cytokine profile of capillary and venous blood of healthy people did not differ significantly. Secondly, our proposed method for determining the subsets of mononuclear cells and capillary blood cytokines profile from the area of psoriatic lesions may be used to monitor local immunity in the patients with psoriasis. This approach is significantly less traumatic than the skin window method and more informative than the studies of venous blood