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1. P-421 Increased myeloid-derived suppressor cells in women with recurrent miscarriage and recurrent implantation failure

Author

N. Subhi-Issa Marin, R. Gil Laborda, E. De la Fuente Muñoz, J. Ochoa Grullón, I Cristóbal, L. Pilar Suárez, M. Calvo Urrutia, M. Fernandez Arquero, and S. Sanchez Ramon

Subjects
Reproductive Medicine, Rehabilitation, Obstetrics and Gynecology
Abstract

Study question We sought to explore the levels of myeloid-derived suppressor cells (MDSCs) in women experiencing recurrent miscarriage (RM) or recurrent implantation failure (RIF) Summary answer MDSCs were significantly expanded in RM patients with respect to fertile healthy women (p = 0.03, 8.25 vs 7.00). What is known already RM and RIF are two distinct disorders that can be associated with a baseline pro-inflammatory state. Different surrogate inflammatory biomarkers have been associated with these disorders in a subgroup of patients, including expanded cytotoxic NK cells, increased pro-inflammatory cytokines or reduced regulatory T cells (TReg). MDSCs are a heterogeneous population of leukocytes, known for their capacity to modulate immune responses. The immunosuppressive function of MDSC in cancer has been well established, while a controversial role in autoimmune diseases has been reported. Typically, there are two major groups of MDSCs in humans: granulocytic/polymorphonuclear MDSCs (PMN-MDSCs) and monocytic MDSCs (M-MDSCs). Study design, size, duration A cross-sectional study to evaluate the levels of MDSCs in women with RM and RIF consecutively referred to our Reproductive Immunology Unit. 55 patients and 23 healthy women were evaluated from 2021 to 2022. RM was defined as the loss of two or more pregnancies and not necessarily consecutive. RIF was defined as the failure to achieve a clinical pregnancy after transferring at least four good-quality embryos. Participants/materials, setting, methods A full fertility screening was performed for all women and their partners as part of routine care. A group of healthy non-pregnant women up to 45 years old with two or more children (proven fertility) and no spontaneous miscarriage was used as control group. Freshly collected blood samples were analyzed by multiparametric flow cytometry using a BD FACS CANTO II. Peripheral blood M-MDSCs were characterized based on HLA-DR, CD14, CD33 and CD45 expression. Main results and the role of chance In this study, we evaluated 55 women, 35 that fulfilled RM criteria and 20 with RIF. In addition, a group of 23 non-pregnant fertile women was included as a control group. We characterized MDSCs as CD14+CD33+CD45+HLA-DR-/LOW, according to previous reports. We observed a significant expansion of peripheral blood MDSCs in RM group compared to the control group (8.25% [6.62-11.9] vs 7.05% [6-7.8], p = 0.03) and RIF group showed a similar increasing trend compared to controls (8.6% [5.65-12.35] vs 7.05% [6-7.8], p = 0.09). We performed ROC curves analysis to evaluate the predictive power. We established an optimal cut-off level of 8.3% in RM group, with a sensitivity of 50% and a specificity of 86.96%. Moreover, women with more than 8.3% had a higher probability of presenting RM compared to the control group (OR: 6.66; CI: 1.68-26.46). MDSCs are characterized by their ability to exert a potent immunosuppressive function. However, recent investigations have shown that MDSCs could act as pro-inflammatory cells under certain inflammatory conditions, such as autoimmune diseases. In this context, MDSCs could worsen the pro-inflammatory state in women, leading to miscarriage or implantation failure. Limitations, reasons for caution In order to validate these results, we should extend our cohort and control group. Moreover, we need to perform a functional evaluation of this subset so we could elucidate their role in these conditions. Wider implications of the findings MDSCs might be a good predictor for RM and RIF patients and with others (as expanded NK cells) may classify a subgroup of patients with a pro-inflammatory profile that can benefit from personalized therapies. Trial registration number Not aplicable

Published
2022
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2. P-367 Centromeric motif AA of the KIR: the best subrogate marker that defines Alloimmune Reproductive Failure

Author

R Gil Laborda, E Carrillo de Albornoz, J Vidal, V Verdú, A De la Fuente, M Fariñas, A Garcia Segovia, D Pintado Vera, J.L Vicario, F Ros Berruezo, F.M Cañadas, M Ordenes, M Fernandez-Arquero, and S Sanchez-Ramon

Subjects
Reproductive Medicine, Rehabilitation, Obstetrics and Gynecology
Abstract

Study question Should the centromeric and telomeric motifs been studied separately and taked them in to account in women with recurrent reproductive failure? Summary answer The results show that when the classic KIR study is performed, there is a 12,5% of AA centromeric KIR patients with reproductive failure unidentified. What is known already During human pregnancy, the decidua is mainly populated by NK lymphocytes that express immunoglobulin-type receptors (KIR) recognizing their HLA-C ligands from trophoblast. The objective of this study was to analyze the association of KIR-HLAC phenotypes in couples with sterility, recurrent pregnancy loss (RPL), recurrent implantation failure (RIF), and fetal death (FD) in comparison with reference population and fertile control. Study design, size, duration It is a retrospective real-life observational study from 2015 to 2019 Participants/materials, setting, methods The frequency of the KIR and HLA-C genes were analyzed in 397 women and their partners with reproductive failure. We compare them with 33 fertile controls and reference Spanish population. KIR and HLA-C genotypes of the patients and controls were determined by Lifecodes Sequence-Specific Oligonucleotides (SSO) Typing KIT (Diagnostica Longwood®) performed at Centro de Transfusiones de la Comunidad de Madrid and those of controls at the Hospital Universitario Clínico San Carlos. Main results and the role of chance Centromeric (Cen) AA was the most frequent KIR genotype in women with unexplained RPL and RIG with respect to reference Spanish population (p = 0.001 and 0.02, respectively). Moreover, cenAA was associated with 1.51-fold risk of RPL and 1.2-fold risk of RIF. On the other hand, BB KIR showed lower risk of any reproductive failure tan non-BB KIR (0.12; p Limitations, reasons for caution The fact of having stratified a lot by groups means that a small number of patients is obtained in each subgroup, which makes it more difficult to conclude the results. Wider implications of the findings Our study demonstrates the importance of the complete KIR genotype study, which would give us more comprehensive information on the behavior of NK cells, moreover we propose that centromeric KIR classification is superior to the KIR classical haplotypes, better identifying those patients with potential KIR-HLA-C mismatch. Trial registration number Not applicable

Published
2022
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4. Early pregnancy

Author

A. Zeadna, H. Holzer, W. Y. Son, E. Demirtas, S. Reinblatt, M. H. Dahan, V. Colleselli, E. D'Costa, L. Wildt, B. Seeber, A. A. Kashevarova, N. A. Skryabin, T. V. Nikitina, I. N. Lebedev, P. P. Bordignon, A. Mugione, V. S. Vanni, P. Vigano, E. Papaleo, M. Candiani, E. Somigliana, G. Amodio, S. Gregori, Y. H. Guo, R. Li, L. L. Wang, S. L. Chen, X. Chen, W. Guo, D. S. Ye, Y. D. Liu, M. M. Renzini, M. Dal Canto, G. Coticchio, R. Comi, C. Brigante, I. Caliari, F. Brambillasca, M. Merola, M. Lain, D. Turchi, G. Karagouga, M. Sottocornola, R. Fadini, M. Z. Wekker, F. Mol, M. van Wely, W. M. Ankum, B. W. Mol, F. van der Veen, P. J. Hajenius, N. M. van Mello, C. Verlengia, E. Alviggi, M. R. Rampini, P. Alfano, I. Pergolini, D. Marconi, N. Iacobelli, M. C. Muzi, G. Gelli, C. Alviggi, A. Colicchia, L. Herraiz-Nicuesa, M. Tejera-Alhambra, A. Garcia-Segovia, R. Ramos-Medina, B. Alonso, J. Gil-Pulido, L. Martin, M. Caballero, M. Rodriguez-Mahou, S. Sanchez-Ramon, P. G. de Jong, S. P. Kaandorp, M. Di Nisio, M. Goddijn, S. Middeldorp, B. Lledo, A. Turienzo, J. A. Ortiz, R. Morales, J. Ten, J. Llacer, R. Bernabeu, J. Gil, J. A. Leon, A. Seyfferth, A. Aguaron, J. Alonso, E. C. de Albornoz, J. Carbone, P. Caballero, E. Fernandez-Cruz, L. Ortiz-Quintana, Y. Y. Lou, F. Jin, Y. M. Zheng, L. J. Li, F. Le, L. Y. Wang, S. Y. Liu, P. P. Pan, C. X. Hu, A. Akoum, A. Bourdiec, R. Shao, C. V. Rao, F. Scarpellini, M. Sbracia, N. Jancar, E. V. Bokal, H. Ban-Frangez, S. Drobnic, S. Korosec, B. Pinter, V. Salamun, M. Yamaguchi, R. Honda, K. Uchino, T. Ohba, H. Katabuchi, O. Leylek, B. Tiras, A. Y. S. E. Saltik, C. Halicigil, N. Kavci, A. Wiser, A. Gilbert, R. Nahum, R. Orvieto, J. Hass, A. Hourvitz, A. Weissman, G. Younes, M. Dirnfeld, A. Hershko, A. Shulma, E. Shalom-Paz, T. Tulandi, S. M. O'Neill, E. Agerbo, L. C. Kenny, T. B. Henriksen, P. M. Kearney, R. A. Greene, P. B. Mortensen, A. S. Khashan, V. S. Talaulikar, B. E. Bax, I. Manyonda, and N. Van Mello

Subjects
Reproductive Medicine, Rehabilitation, Obstetrics and Gynecology
Published
2013
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5. Typhim vi immunization assists to discriminate primary antibody responses in hematological malignancies

Author

J. Ochoa-Grullón, C. Orte, A. Rodríguez de la Peña, K. Guevara-Hoyer, G. Cordero Torres, M. Fernández-Arquero, I. Serrano-García, M.J. Recio, R. Pérez de Diego, and S. Sánchez-Ramón

Subjects
Primary responses, secondary responses, specific polysaccharide Ab response, Pneumo 23, Typhim Vi, Hematological Malignancies, Science
Abstract

ABSTRACT: Assessment of specific antibody (Ab) production to polysaccharide antigens is clinically relevant, identifying patients at risk for infection by encapsulated bacteria and thus enabling a more rigorous selection of patients that can benefit of immunoglobulin replacement therapy. Classically, the gold-standard test is the measurement of antibody production to pure polysaccharide pneumococcal (PPV) immunization. Several factors, including introduction of conjugate vaccination schedule, serotyping analysis, high baseline Ab levels, have hindered the evaluation of polysaccharide antigens. This is even more difficult in secondary immunodeficiencies (SID), where patients can show secondary responses despite lack of primary antibody responses and present with recurrent or severe infections. Assessment of specific Ab production to pure Salmonella typhi Vi polysaccharide (TV) immunization has been proposed as a complementary test to PPV, given its low seroprevalence. To set the optimal cut-off value for PPV and TV response in SID, we tested different biostatistical methodologies, including ROC analysis, Youden index, Union index and Closest-topleft in a cohort of 42 SID patients and 24 healthy controls. The statistically chosen cut-offs value pre-post TV Ab ratio was ≥5, (sensitivity of 90%, specificity of 100%) and a postvaccination TV concentration of 28.5 U/mL (sensitivity of 90%, specificity of 95%), showing relevant clinical correlate.

Published
2020
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6. Cambios en las Subpoblaciones de Linfocitos B y T en el Título de Anticuerpos Anti-Acuaporina-4 tras el Tratamiento de un Brote Agudo con Inmunoglobulinas y Rituximab

Author

C. de Andrés, R. Teijeiro, A. Saiz, P. Fernández, and S. Sánchez-Ramón

Subjects
Neurology. Diseases of the nervous system, RC346-429
Abstract

Resumen: Antecedentes: La neuromielitis óptica (NMO) es una enfermedad predominantemente humoral mediada por anticuerpos IgG-NMO/AQP-4. Sin embargo, no se conoce bien la contribución de las diferentes subpoblaciones de células B y T en su patogenia o en la respuesta a los tratamientos. Objetivos: Describir los cambios clínicos e inmunológicos asociados al tratamiento con inmunoglobulinas intravenosas (Ig-IV) y rituximab (RTX) en una paciente con un brote grave de NMO y síntesis intratecal de IgG-NMO/AQP-4 que no había respondido a metilprednisolona y recambio plasmático. Métodos: Se analizaron, de forma secuencial en el LCR y en la sangre periférica (SP), las subpoblaciones linfocitarias mediante citometría de flujo multiparamétrica y los IgG-NMO/AQP-4, antes y después del tratamiento con Ig-IV y RTX. Resultados: En el LCR antes del tratamiento, y comparado con la SP, predominaban las células T CD4+ y estaban menos representadas las T CD8+ y las B CD19+. Tras el tratamiento, el porcentaje de células T CD4+ se mantuvo alto, el de T CD8+ aumentó y el de B CD19+ disminuyó, aunque menos que en la SP. Al comparar los LCR se vio que tras la terapia el porcentaje de células T CD8+ memoria efectoras y efectoras había aumentado, y el de células B memoria IgM y el de células B con cambio de isotipos, disminuido. Los cambios observados fueron paralelos a la negativización de los IgG-NMO/AQP-4 y a la mejoría clínica. Conclusiones: Nuestros hallazgos confirman que, además de una respuesta inmunitaria humoral intratecal durante el brote de NMO, subpoblaciones específicas de células B y T participan en la modulación de la inflamación dentro y fuera del sistema nervioso central. Abstract: Background: There is increasing evidence supporting that neuromyelitis optica (NMO) is an inflammatory humoral mediated disorder associated with NMO-IgG/AQP-4 antibodies. However, little is known about the subsets of B cells and T cells that contribute to the pathogenesis or therapy response. Objectives: To describe the clinical and immunological changes associated with intravenous immunoglobulins (IV-Igs) plus rituximab (RTX) in a patient with a severe acute attack of NMO and intrathecal synthesis of NMO-IgG/AQP-4, who previously did not respond to intravenous methylprednisolone and plasma exchange. Methods: We sequentially analysed the levels of NMO-IgG/AQP-4 by immunohistochemistry, and B and T cells subsets by multiparametric flow-cytometry, in the CSF and peripheral blood (PB), before and alter IV-Igs plus RTX therapy. Results: In the CSF before treatment, and compared with PB, there was a higher percentage of CD4+ T cells and a lower percentage of CD8+ T cells and CD19+ B cells. After therapy, the percentage of CD4+ T cells remained high, and that of CD8+ T cells increased. The observed decrease in the percentage of CD19+ B cells was lower than in the PB. When the CSF was compared, it was found that the percentage of effector-memory and effector CD8+ T cells had increased after therapy, and that of IgM memory B cells and switched-memory B cells decreased. The observed changes paralleled the decrease of NMO-IgG/AQP-4 results to negative and the clinical improvement. Conclusions: Our findings confirm that, besides intrathecal humoral immune response against AQP4, B and T cell subsets are involved in the modulation of inflammation within and outside the central nervous system. Palabras clave: Neuromielitis óptica, Líquido cefalorraquídeo, Autoanticuerpos Anti-acuaporina-4, Subclases de células B CD19+ y T CD8+, Keywords: Neuromyelitis optica, Cerebrospinal fluid, Anti-aquaporin-4 autoantibodies, CD19+ and CD8+ T cell subsets

Published
2015
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7. Changes in B and T-cell subsets and NMO-IgG/AQP-4 levels after immunoglobulins and rituximab treatment for an acute attack of neuromyelitis optica

Author

C. de Andrés, R. Teijeiro, A. Saiz, P. Fernández, and S. Sánchez-Ramón

Subjects
Neurology. Diseases of the nervous system, RC346-429
Abstract

Background: There is increasing evidence supporting that neuromyelitis optica (NMO) is an inflammatory humoral mediated disorder associated with NMO-IgG/AQP-4 antibodies. However, little is known about the subsets of B cells and T cells that contribute to the pathogenesis or therapy response. Objectives: To describe the clinical and immunological changes associated with intravenous immunoglobulins (IV-Igs) plus rituximab (RTX) in a patient with a severe acute attack of NMO and intrathecal synthesis of NMO-IgG/AQP-4, who previously did not respond to intravenous methylprednisolone and plasma exchange. Methods: We sequentially analysed the levels of NMO-IgG/AQP-4 by immunohistochemistry, and B and T cells subsets by multiparametric flow-cytometry, in the CSF and peripheral blood (PB), before and after IV-Igs plus RTX therapy. Results: In the CSF before treatment, and compared to PB, there was a higher percentage of CD4+ T cells and a lower percentage of CD8+ T cells and CD19+ B cells. After therapy, the percentage of CD4+ T cells remained high, and that of CD8+ T cells increased. The observed decrease in the percentage of CD19+ B cells was lower than in the PB. When the CSF was compared, it was found that the percentage of effector-memory and effector CD8+ T cells had increased after therapy, and that of IgM memory B cells and switched-memory B cells decreased. The observed changes paralleled the decrease of NMO-IgG/AQP-4 results to negative and the clinical improvement. Conclusions: Our findings confirm that, besides intrathecal humoral immune response against AQP4, B and T cell subsets are involved in the modulation of inflammation within and outside the central nervous system. Resumen: Antecedentes: La neuromielitis óptica (NMO) es una enfermedad predominantemente humoral mediada por anticuerpos IgG-NMO/AQP-4. Sin embargo, no se conoce bien la contribución de las diferentes subpoblaciones de células B y T en su patogenia o en la respuesta a los tratamientos. Objetivos: Describir los cambios clínicos e inmunológicos asociados al tratamiento con inmunoglobulinas intravenosas (Ig-IV) y rituximab (RTX) en una paciente con un brote grave de NMO y síntesis intratecal de IgG-NMO/AQP-4 que no había respondido a metilprednisolona y recambio plasmático. Métodos: Se analizaron, de forma secuencial en el LCR y en la sangre periférica (SP), las subpoblaciones linfocitarias mediante citometría de flujo multiparamétrica y los IgG-NMO/AQP-4, antes y después del tratamiento con Ig-IV y RTX. Resultados: En el LCR antes del tratamiento, y comparado con la SP, predominaban las células T CD4+ y estaban menos representadas las T CD8+ y las B CD19+. Tras el tratamiento, el porcentaje de células T CD4+ se mantuvo alto, el de T CD8+ aumentó y el de B CD19+ disminuyó, aunque menos que en la SP. Al comparar los LCR se vio que tras la terapia el porcentaje de células T CD8+ memoria efectoras y efectoras había aumentado, y el de células B memoria IgM y el de células B con cambio de isotipos, disminuido. Los cambios observados fueron paralelos a la negativización de los IgG-NMO/AQP-4 y a la mejoría clínica. Conclusiones: Nuestros hallazgos confirman que, además de una respuesta inmunitaria humoral intratecal durante el brote de NMO, subpoblaciones específicas de células B y T participan en la modulación de la inflamación dentro y fuera del sistema nervioso central. Keywords: Neuromyelitis optica, Cerebrospinal fluid, Anti-aquaporin-4 autoantibodies, CD19+ and CD8+ T cell subsets, Palabras clave: Neuromielitis óptica, Líquido cefalorraquídeo, Autoanticuerpos Anti-acuaporina-4, Subclases de células B CD19+ y T CD8+

Published
2015
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