A growing body of evidence has demonstrated that soy isoflavone consumption may protect against the development of various chronic diseases. This protection could be linked to isoflavone-induced alterations in immune function. However, recent in vitro and animal studies suggest that soy isoflavones may either enhance or suppress immunocompetence, depending upon the isoflavone concentration, target tissue, and a number of other factors. To date, no study has investigated the effect of dietary soy isoflavone supplementation on immune parameters in humans. Therefore, the purpose of this double-blind, placebo-controlled, 4 wk intervention trial was to investigate whether supplementation with soy isoflavones alters indices of immune function in postmenopausal women. Twenty healthy women (50-69 yr), who were not on hormone replacement therapy, were randomly divided into 2 treatment groups. The supplemented group (n=10) consumed soy isoflavone tablets (100 mg/d) for 4 wk, while the control group (n=10) received placebo tablets. Fasting blood samples were drawn at baseline and on d 28 to assess specific immune parameters. In addition, plasma concentrations of genistein and daidzein were quantified at baseline and at the end of the intervention period. Despite high individual variability among subjects, there was a significant increase (p<0.005) in plasma isoflavone concentration in the supplemented group. However, all assessed immune parameters remained unchanged after supplementation and did not differ between the 2 treatment groups. In conclusion, this study suggests that short-term soy isoflavone supplementation at physiologically attainable concentrations does not alter the aforementioned immune parameters in healthy postmenopausal women. Due to the conflicting data concerning the effect of dietary soy isoflavones on immune function, further research in this area is warranted. / Master of Science

A growing body of evidence has demonstrated that soy isoflavone consumption may protect against the development of various chronic diseases. This defense could be linked to isoflavone-induced alterations in immune function. However, to date, no study has examined the effect of soy isoflavone supplementation on human immunity in vivo. Establishing whether isoflavones affect immunity in aging adults is particularly relevant since compromised immune function has been observed in this population. Therefore, the purpose of this double-blind, placebo-controlled, 4-wk intervention trial was to investigate whether supplementation with soy isoflavones influenced the distribution and/or function of specific lymphocytes in postmenopausal women. Healthy postmenopausal women (50-69 y), who were not using hormone replacement therapy, were randomly divided into 2 treatment groups. The experimental group (n=9) consumed two-50 mg soy isoflavone tablets/d for 4 wk, while the control group (n=9) received placebo tablets. Fasting blood samples were drawn at baseline and on d 28 to assess distribution of T-helper cells (CD3+CD4+), T-cytotoxic cells (CD3+CD8+), total T lymphocytes (CD3+), B lymphocytes (CD19+) and natural killer (NK) cells (CD16+CD56+) via flow cytometry. Cytotoxicity of NK cells was quantified based on lactate dehydrogenase release of lysed K562 cancer cells following co-culture with NK cells from subjects. Analysis of plasma isoflavone concentrations by HPLC demonstrated a significant increase (p<0.005) in plasma genistein concentration in the experimental group after 4 wk of supplementation. However, there was no alteration in lymphocyte distribution or NK cell activity in response to isoflavone supplementation, suggesting that short-term soy isoflavone supplementation does not alter these parameters of immunity in healthy postmenopausal women. / Master of Science

Two important mechanisms of lymphocyte-mediated cytotoxicity, one perforin based and the other Fas ligand (FasL) based, have been characterized recently. It has also been shown that CD44, an adhesion molecule, can participate in signaling cytotoxic activity of cytotoxic T lymphocytes (CTLs). In the current study we tested the hypothesis that activation of natural killer (NK) or lymphokine activated killer (LAK) cells induces the expression of FasL, perforin, and CD44 which together contribute towards increased cytolytic activity. To this effect, we used wild-type mice, perforin-knockout mice, and mice lacking a functional FasL. We observed that both interleukin-2 (IL-2) and Poly I:C triggered NK/LAK cells to lyse targets through the perforin- and FasL- pathways. In addition, Fas+ tumor targets were more susceptible to lysis by poly I:C and IL-2 activated NK/LAK cells when compared to Fas- targets. Furthermore, Fas- tumor cells injected subcutaneously into syngeneic mice could grow and induce tumors, whereas, Fas+ tumors were rejected. IL-2 treatment increased the CD44 expression on NK cells, which was responsible for the lysis of endothelial cells through its ligand, hyaluronate. Upregulation of perforin and FasL in activated NK/LAK cells may explain why such cells can kill a wide variety of tumor cells efficiently. On the other hand, activated NK/LAK cells express increase increased levels of CD44 and use this molecule to mediate cytotoxicity of endothelial cells, which may account for the vascular leak seen during IL-2 therapy. / Master of Science

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 4 uppsatser.

Les dernières avancées dans les traitements des hémopathies aboutissent à un meilleurs taux de rémission complète ainsi qu' à de meilleurs taux de survie après traitement. Cependant les risques de rechutes restent élevés. Notre projet s'inscrit dans la compréhension du rôle des cellules NK dans l'évolution de ce type de pathologies. Dans une première partie nous nous sommes intéressés à la polyglobulie de Vaquez. Cette pathologie présente une évolution lente et progressive, et elle est caractérisée par une mutation de JAK2 présente dans la lignée myéloïde chez plus de 95% des patients. Nous avons cherché à détecter la mutation dans les cellules NK de patients, puis, pour savoir si la mutation avait un effet sur les NK, nous avons exploré leurs fonctions in vitro. Nos résultats ont montré que, bien que la mutation soit présente dans les cellules NK, elle ne semble pas avoir d'impact sur les fonctions des cellules NK que nous avons pu tester. Nous en avons conclu que l'évolution de la polyglobulie de Vaquez en leucémie n'était peut-être pas due à une perte de fonction des NK mais plutôt à leur inhibition par l'environnement cellulaire.Dans une deuxième partie nous avons étudié la régulation des natural cytotoxicity receptors dans la leucémie aiguë myéloïde. D'apres des travaux antérieurs nous avons émis l'hypothèse que l'expression des trois NCR aurait une régulation commune s'effectuant au niveau de transcription de leurs gènes. Nos recherches bio-informatiques ainsi que notre expérimentation d'immunoprécipitation de la chromatine (Chip) montrent que le facteur de transcription ETS-1 semble être impliqué dans la régulation commune aux trois NCR. / The latest advances in blood disorders treatments lead to a better complete remission rate and a better survival rate after treatment. However, the risk of relapse remains high. Our project is included in the understanding of NK cells role in the development of these diseases.In a first part, we focused on polycythemia Vera for several reasons: the pathology has a slowly progressive disease, and it is characterized by the presence of JAK2 mutation for > 95% patients. We wanted to know if this mutation was found in NK cells from PV patients and what effects the mutation had on NK cells functions. Our results have shown that although the mutation was found in NK cells, it appears to have no impact on NK cells functions. We conclude that the evolution of PV to leukemia is not due to a loss of NK cell functions but to their inhibition by cellular environment.In a second part, we investigated the regulation of natural cytotoxicity receptors in acute myeloid leukemia because previous works have shown that NCR are weakly expressed in AML patients, that this down-regulation is acquired during evolution of AML and reversible after complete remission, ant that NCR weak expression is related to poor prognosis. We supposed that the expression of the three NCR has a common regulation at genes transcription level. Our bioinformatic researches and our experiment of chromatin immunoprecipitation show that ETS-1 transcription factor is a good candidate involved in the common regulation of the three NCR.

Early pregnancy is characterized by complex interactions between blood vessels, leukocytes, and conceptus-derived trophoblasts within the gestational uterus. Uterine Natural Killer (uNK) cells become the most abundant leukocyte during decidualization and produce a wide array of angiogenic factors, yet little is known regarding their early pregnancy functions. To characterize the role(s) of uNK cells, whole mount in situ immunohistochemistry of live early implant sites was performed. A timecourse examination of murine early pregnancy (virgin, and gd4.5-9.5) implantation sites was performed. Comparison of Gd6.5, 8.5 and 9.5 implant sites from BALB/c+/+ controls (BALB/c) and BALB/c-Rag2-/-Il2rg-/- (alymphoid) identified anomalies that result from the absence of lymphocytes. In alymphoid decidua basalis, mesometrial angiogenesis was widespread but pruning of nascent vessels within alymphoid decidua basalis was deficient. As early gestation progressed, vessels of alymphoid decidua basalis showed no evidence for remodeling. Alymphoid implantation sites showed ~24h delay in uterine lumen closure and embryonic development. To determine if uNK cells would normalize the anomalies observed in alymphoid implantation sites, adoptive cell transfer of NK+ B- T- marrow to alymphoid mice was performed. All of the above anomalies were reversed by adoptive transfer of NK+B-T- marrow. My results suggest that uNK cells support vascular growth and development which ensures the decidua can support the growing conceptus early in pregnancy prior to formation and function of the placenta. Human decidual NK cells may fill similar roles and be important targets for strategies designed to correct intra-uterine growth restriction. / Thesis (Master, Anatomy & Cell Biology) -- Queen's University, 2013-08-02 08:42:06.487

Embora a obesidade esteja associada à disfunção imune, com incidência aumentada de infecções e alguns tipos de cânceres, há poucos estudos que avaliaram parâmetros imunológicos em pacientes obesos graves. Além disso, há um número limitado de trabalhos analisando o efeito da perda de peso sobre parâmetros imunológicos na obesidade grave. Desta forma, o objetivo do presente trabalho foi avaliar a influência da perda de peso de pacientes com obesidade grau III submetidos à cirurgia de derivação gastrojejunal em Y de Roux (DGJYR) em parâmetros imunológicos. A produção de citocinas associadas com a resposta imune adquirida (IL-2, IL-4, IL-10 e IFN-) e inata (TNF- e IL-6) por células mononucleares de sangue periférico (PBMC), o perfil das populações de linfócitos e a atividade citotóxica de células natural killer (NK), além de citocinas associadas a sua função e desenvolvimento (IL-12 e IL-18), foram avaliados em vinte e oito pacientes não diabéticos, sedentários, com obesidade grau III (20 mulheres e 8 homens, com média de idade de 39,9 ± 10,9 anos e IMC de 49,5 ± 7,1kg/m2) no pré-operatório e 6 meses após a cirurgia. As PBMC foram estimuladas com o mitógeno fitohemaglutinina (PHA) e as citocinas produzidas foram quantificadas por ELISA. O perfil das populações de linfócitos foi avaliado por citometria de fluxo. A citotoxicidade mediada por células NK foi determinada pelo ensaio de liberação de LDH por células alvo K562. A perda de peso foi de 35,3 ± 4,5 kg, com uma significativa redução no IMC seis meses após a cirurgia (-12,9 ± 0,9 kg/m2, p< 0,001). Nenhuma das populações de linfócitos analisadas apresentou modificação no 6º mês após a cirurgia. Observou-se aumento significativo da proliferação de linfócitos seis meses após a cirurgia (p= 0,0026). Houve aumento pósoperatório nas concentrações de IFN-, IL-12 e IL-18 produzidas por PBMC após estímulo com PHA, enquanto a IL-2 apresentou uma tendência ao aumento (p= 0,07). As demais citocinas não apresentaram variação significativa. A atividade citotóxica das células NK aumentou seis meses após a cirurgia [17,1 ± 14,7% no pré vs 51,8 ± 11,3% 6 meses pósoperatório, na proporção 40:1 (célula NK:célula alvo); p< 0,001], mostrando recuperação quando se compara aos valores obtidos em indivíduos controle, pareados por idade e sexo, de peso normal [proporção 40:1 (célula NK:célula alvo) de 45,4 ± 7,8%]. Houve aumento de atividade citotóxica em todos os pontos da curva no pós-operatório em cerca de 79% da amostra (22 pacientes). Os resultados obtidos demonstram que a perda de peso induzida por DGJYR aumenta a produção de algumas citocinas relacionadas com a função das células NK e melhora a sua atividade citotóxica. As alterações na função de células NK e do nível de citocinas envolvidas com a atividade destas células podem explicar a propensão ao desenvolvimento de infecções e cânceres associados com a obesidade. Os dados obtidos neste estudo sugerem que a cirurgia bariátrica pode ter impacto positivo sobre estes fatores. / Although obesity is related to immune dysfunction, with a higher incidence of infections and some types of cancer, few studies have evaluated immunological parameters in severely obese patients. Moreover, a limited set of studies have analyzed the effect of weight loss in immunological parameters in severely obese patients. Thus, the objective of this thesis was to evaluate the influence of weight loss induced by Roux en-Y gastric bypass in patients with grade III obesity in immunological parameters. The production of cytokines associated with acquired (IL-2, IL-4, IL-10 and IFN-) and innate (TNF- e IL-6) immune responses from peripheral blood mononuclear cells (PBMCs), the profile of lymphocytes populations and the cytotoxic activity of natural killer cells (NK), besides cytokines related with NK cell cytotoxic function and development (IL-12 e IL-18), were analyzed in 28 non-diabetic and sedentary patients with grade III obesity (20 women and 8 men, 39,9 ± 10,9 years and BMI 49,5 ± 7,1 kg/m2) before and 6 months after RYGB. PBMCs were stimulated with the mitogen phytohemagglutinin (PHA) and cytokines were measure by ELISA. The profile of lymphocytes populations was evaluated by flow cytometry. NK cell cytotoxicity was determined by the lactate dehydrogenase release assay from K562 lysed target cells. The weight loss 6 months after surgery was 35.3±4.5 kg and there was a significant post-surgical decrease in BMI at this point (-12.9±0.9 kg/m2, p<0.001). No significant differences were found in the lymphocytes populations after surgery. It was observed a significant increase in the lymphocytes proliferation six months after surgery (p= 0.0026). There was also a post-surgical increase in the production of IFN-, IL-12 e IL-18 from PBMC stimulated with PHA, while there was a trend towards the increase of the IL-2 production (p=0.07). The other cytokines analyzed were not altered. Cytotoxic activity of NK cells was significantly enhanced 6 months after RYGB [17.1±14.7% before RYGB vs 51.8±11.3% at 6 months after, at effector to target cell (NK cell:K562 cell) ratio 40:1; p<0.001], and it was in the same range when compared to data obtained from controls with normal BMI matched for age and gender (45,4 ± 7,8% at NK cell:K562 cell ratio 40:1). There was a significant post-surgical improvement in all points of the cytotoxic activity curve in almost 79% of the sample (22 patients). In conclusion, the data obtained show that the weight loss induced by RYGB increases the production of cytokines related with NK cell cytotoxic function and improves its activity. The impairment in NK cells cytotoxic activity and cytokines observed in patients with severe obesity may explain their propensity to develop infections and cancer. Our data suggests that the weight loss induced by bariatric surgery can positively impact these factors.

Objetivo: o objetivo deste estudo foi avaliar a prevalência de autoanticorpos e a dosagem da concentração de células Natural Killer (NK) no sangue periférico em pacientes com endometriose. Métodos: Entre dezembro de 2004 e dezembro de 2007 foram avaliadas 155 pacientes submetidas a videolaparoscopia, divididas em um grupo sem endometriose(n=55) e outro com endometriose (n=100). Foi coletada amostra de sangue periférico de todas as pacientes no momento da laparoscopia e nessa amostra foi realizada a quantificação do percentual de células NK em relação aos linfócitos periféricos (por citometria de fluxo), e a determinação dos seguintes auto-anticorpos: anticorpos antinucleares (ANA, por imunofluorescência indireta), anticorpos antitireoglobulina e antiperoxidase (anti-TG e anti-TPO, por eletroquimioluminescência), anticorpos anticardilipina e antifosfatidilserina (aCL e aPS IgG, IgM e IgA, todos por ensaio imunoenzimático). Além da presença de endometriose, essas pacientes também foram avaliadas quanto ao estadiamento, os locais de doença, relações com a fase do ciclo, e a classificação histológica dessa doença. Resultados: as pacientes com endometriose apresentaram percentual de células NK (média DP de 15,3 9,8%) superiores àquelas sem a doença (média DP de 10,6 5,8%), p<0,001. Quanto aos autoanticorpos, as portadoras de endometriose também apresentaram positividade para ANA mais frequentemente (33%) que as pacientes do grupo controle (12,7%), p=0,006. Quanto aos anti-TG, anti-TPO, anti-CL (IgG, IgM e IgA) e aPS ( IgG, IgM e IgA), não houve diferenças estatísticas quanto à sua positividade. As células NK também mostraram-se mais elevadas nas protadoras de endometriose em estádios avançados e naquelas com comprometimento de retossigmóide, grupo no qual encontramos o maior percentual de células NK com concentração média de 19,8 10,3%. Concentrações de células NK 12,5% podem ser usadas como marcadores de endometriose em retossigmóide, com sensibilidade de 73% e especificidade de 65%. Utilizando-se de um modelo estatístico de probabilidades, demonstramos que associação desse marcador (NK 12,5%) com a presença de sintomas como dor e/ou sangramento intestinal durante a menstruação nos possibilitou estimar uma probabilidade de comprometimento de retossigmóide de 60,4%. Conclusões: pacientes com endometriose apresentam maior concentração de células NK periféricas, além de maior prevalência de ANA positivo em relação àquelas sem endometriose. As células NK aumentam nas pacientes com endometriose predominantemente nos estádios avançados, com comprometimento de retossigmóide. Nesse sentido poderiam ser utilizadas como marcadores diagnósticos desse tipo de comprometimento da doença, principalmente se forem avaliadas em conjunto com os sintomas das pacientes / Objectives: The objective of this study was to evaluate the prevalence of autoantibodies and the percentage of natural killer (NK) cells in the peripheral blood of patients with endometriosis. Methods: Between December 2004 and December 2007, 155 patients submitted to videolaparoscopy were evaluated. Patients were divided into two groups: one group of women without endometriosis (n = 55) and another in which all the women had endometriosis (n = 100). Samples of peripheral blood were collected from all the patients at the time of laparoscopy and flow cytometry was used to determine the percentage of NK cells in relation to peripheral blood lymphocytes in these samples. In addition, the following autoantibodies were measured: antinuclear antibodies (ANA) by indirect immunofluorescence, anti-thyroglobulin and anti-thyroid peroxidase antibodies (anti-TG and anti-TPO) by electrochemiluminescence, and anticardiolipin and anti-phosphatidylserine antibodies (aCL and aPS IgG, IgM and IgA), all performed using immunoenzymatic assay. In addition to the presence of endometriosis, these patients were also evaluated with respect to staging, to the sites of the disease, any association with the phase of the menstrual cycle and the histological classification of the disease. Results: The patients with endometriosis had a higher percentage of NK cells (15.3 ± 9.8%; mean ± SD) compared to those without the disease (10.6 ± 5.8%; mean ± SD), (p<0.001). Evaluation of the autoantibodies showed that positivity for ANA was more common in the group of patients with endometriosis (33%) compared to the patients in the control group (12.7%), (p = 0.006). With respect to anti-TG, anti-TPO, aCL (IgG, IgM and IgA) and aPS (IgG, IgM and IgA), no statistically significant differences were found between the groups of patients with or without endometriosis. NK cell concentrations were also found to be higher in patients with advanced stages of endometriosis and in those in whom the rectosigmoid was affected by the disease, this being the group in which the highest percentage of NK cells was found, with mean concentrations of 19.8 ± 10.3%. NK cell concentrations 12.5% may be used as markers of endometriosis of the rectosigmoid, with sensitivity of 73% and specificity of 65%. Using a statistical model of probability, these findings showed that the association of this marker (NK 12.5%) with the presence of symptoms such as pain and/or intestinal bleeding during menstruation permitted an estimation to be made of a likelihood of 60.4% of rectosigmoid endometriosis. Conclusions: Patients with endometriosis have higher percentages of peripheral NK cells, as well as a greater prevalence of positive ANA compared to those without endometriosis. The concentration of peripheral NK cells increases in patients with endometriosis, predominantly in patients with advanced stages of the disease and those in whom the rectosigmoid is affected. Therefore, the concentration of NK cells in peripheral blood could be used as a diagnostic marker of this type of endometriosis, particularly when evaluated together with patients symptoms

Objetivo: o objetivo deste estudo foi avaliar a prevalência de autoanticorpos e a dosagem da concentração de células Natural Killer (NK) no sangue periférico em pacientes com endometriose. Métodos: Entre dezembro de 2004 e dezembro de 2007 foram avaliadas 155 pacientes submetidas a videolaparoscopia, divididas em um grupo sem endometriose(n=55) e outro com endometriose (n=100). Foi coletada amostra de sangue periférico de todas as pacientes no momento da laparoscopia e nessa amostra foi realizada a quantificação do percentual de células NK em relação aos linfócitos periféricos (por citometria de fluxo), e a determinação dos seguintes auto-anticorpos: anticorpos antinucleares (ANA, por imunofluorescência indireta), anticorpos antitireoglobulina e antiperoxidase (anti-TG e anti-TPO, por eletroquimioluminescência), anticorpos anticardilipina e antifosfatidilserina (aCL e aPS IgG, IgM e IgA, todos por ensaio imunoenzimático). Além da presença de endometriose, essas pacientes também foram avaliadas quanto ao estadiamento, os locais de doença, relações com a fase do ciclo, e a classificação histológica dessa doença. Resultados: as pacientes com endometriose apresentaram percentual de células NK (média DP de 15,3 9,8%) superiores àquelas sem a doença (média DP de 10,6 5,8%), p<0,001. Quanto aos autoanticorpos, as portadoras de endometriose também apresentaram positividade para ANA mais frequentemente (33%) que as pacientes do grupo controle (12,7%), p=0,006. Quanto aos anti-TG, anti-TPO, anti-CL (IgG, IgM e IgA) e aPS ( IgG, IgM e IgA), não houve diferenças estatísticas quanto à sua positividade. As células NK também mostraram-se mais elevadas nas protadoras de endometriose em estádios avançados e naquelas com comprometimento de retossigmóide, grupo no qual encontramos o maior percentual de células NK com concentração média de 19,8 10,3%. Concentrações de células NK 12,5% podem ser usadas como marcadores de endometriose em retossigmóide, com sensibilidade de 73% e especificidade de 65%. Utilizando-se de um modelo estatístico de probabilidades, demonstramos que associação desse marcador (NK 12,5%) com a presença de sintomas como dor e/ou sangramento intestinal durante a menstruação nos possibilitou estimar uma probabilidade de comprometimento de retossigmóide de 60,4%. Conclusões: pacientes com endometriose apresentam maior concentração de células NK periféricas, além de maior prevalência de ANA positivo em relação àquelas sem endometriose. As células NK aumentam nas pacientes com endometriose predominantemente nos estádios avançados, com comprometimento de retossigmóide. Nesse sentido poderiam ser utilizadas como marcadores diagnósticos desse tipo de comprometimento da doença, principalmente se forem avaliadas em conjunto com os sintomas das pacientes / Objectives: The objective of this study was to evaluate the prevalence of autoantibodies and the percentage of natural killer (NK) cells in the peripheral blood of patients with endometriosis. Methods: Between December 2004 and December 2007, 155 patients submitted to videolaparoscopy were evaluated. Patients were divided into two groups: one group of women without endometriosis (n = 55) and another in which all the women had endometriosis (n = 100). Samples of peripheral blood were collected from all the patients at the time of laparoscopy and flow cytometry was used to determine the percentage of NK cells in relation to peripheral blood lymphocytes in these samples. In addition, the following autoantibodies were measured: antinuclear antibodies (ANA) by indirect immunofluorescence, anti-thyroglobulin and anti-thyroid peroxidase antibodies (anti-TG and anti-TPO) by electrochemiluminescence, and anticardiolipin and anti-phosphatidylserine antibodies (aCL and aPS IgG, IgM and IgA), all performed using immunoenzymatic assay. In addition to the presence of endometriosis, these patients were also evaluated with respect to staging, to the sites of the disease, any association with the phase of the menstrual cycle and the histological classification of the disease. Results: The patients with endometriosis had a higher percentage of NK cells (15.3 ± 9.8%; mean ± SD) compared to those without the disease (10.6 ± 5.8%; mean ± SD), (p<0.001). Evaluation of the autoantibodies showed that positivity for ANA was more common in the group of patients with endometriosis (33%) compared to the patients in the control group (12.7%), (p = 0.006). With respect to anti-TG, anti-TPO, aCL (IgG, IgM and IgA) and aPS (IgG, IgM and IgA), no statistically significant differences were found between the groups of patients with or without endometriosis. NK cell concentrations were also found to be higher in patients with advanced stages of endometriosis and in those in whom the rectosigmoid was affected by the disease, this being the group in which the highest percentage of NK cells was found, with mean concentrations of 19.8 ± 10.3%. NK cell concentrations 12.5% may be used as markers of endometriosis of the rectosigmoid, with sensitivity of 73% and specificity of 65%. Using a statistical model of probability, these findings showed that the association of this marker (NK 12.5%) with the presence of symptoms such as pain and/or intestinal bleeding during menstruation permitted an estimation to be made of a likelihood of 60.4% of rectosigmoid endometriosis. Conclusions: Patients with endometriosis have higher percentages of peripheral NK cells, as well as a greater prevalence of positive ANA compared to those without endometriosis. The concentration of peripheral NK cells increases in patients with endometriosis, predominantly in patients with advanced stages of the disease and those in whom the rectosigmoid is affected. Therefore, the concentration of NK cells in peripheral blood could be used as a diagnostic marker of this type of endometriosis, particularly when evaluated together with patients symptoms

Lytic effector function of Natural Killer (NK) cells and CD8+ T cells occurs through discrete and regulated cell biological steps triggered by recognition of diseased cells. Recent studies of the NK cell synapse support the idea that dynamic nanoscale rearrangements in cortical filamentous (F)-actin are a critical cell biological checkpoint for lytic granule access to NK cell membrane. Loss of function mutations in the LYST gene, a well-characterised cause of Chediak- Hegashi syndrome (CHS), result in the formation of giant lysosomal organelles including lytic granules. Here, we report a mismatch between the extent of cortical F-actin remodelling and enlarged lytic granules that limits the functionality of LYST- deficient NK cells in a human model of CHS. Using super-resolution stimulated emission depletion (STED) microscopy we found that LYST-deficient NK cells had nanoscale rearrangements in the organisation of cortical actin filaments that were indistinguishable from control cells- despite a 2.5-fold increase in the size of polarised granules. Importantly, treatment of LYST-deficient NK cells with actin depolymerising drugs increased the formation of small secretory domains at the synapse and restored their ability to lyse target cells. These data establish that sub-synaptic F-actin is the major factor limiting the release of enlarged lytic granules from CHS NK cells, and reveal a novel target for therapeutic interventions. While the importance of cortical actin filaments in NK cell cytotoxicity have been established, its persistence at the early stages of T cell synapse formation is disputed. We studied the organisation of cortical actin filaments in synapses formed by primary human T cells using STED microscopy and detected intact cortical actin filaments in key T cell effector subsets including memory CD8+ T cells as early as 5-minutes post-activation. Quantitative analysis revealed that activation specific rearrangements in cortical actin filaments at both CD4+ and CD8+ T cell synapses serve to increase the space between filaments. Additionally, comparison of cytolytic T cells with freshly isolated and IL-2 activated primary NK cells revealed that rapid maturation of the cortical actin meshwork is a specific feature of CD8+ T cell lytic synapses. Using chemical inhibition of actin nucleators, we show that increased cortical relaxation is mediated primarily by the activity of actin related proteins (Arp) -2/3. Taken together, these data establish the critical requirement for dynamic rearrangements in cortical actin filaments at lytic synapses but underscore cell-specific differences in its regulation.