Tumor cells and cancer-associated fibroblasts: A synergistic crosstalk to promote thyroid cancer

Thyroid cancer is the most common endocrine malignancy. Although most thyroid cancer patients are successfully treated and have an excellent prognosis, a percentage of these patients will develop aggressive disease and, eventually, progress to anaplastic thyroid cancer. Since most patients with this type of aggressive thyroid carcinoma will die from the disease, new treatment strategies are urgently needed. Tumor cells live in a complex and dynamic tumor microenvironment composed of different types of stromal cells. Cancer-associated fibroblasts (CAFs) are one of the most important cell components in the tumor microenvironment of most solid tumors, including thyroid cancer. CAFs originate mainly from mesenchymal cells and resident fibroblasts that are activated and reprogrammed in response to paracrine factors and cytokines produced and released by tumor cells. Upon reprogramming, which is distinguished by the expression of different marker proteins, CAFs synthesize and secret soluble factors. The secretome of CAFs directly impacts different functions of tumor cells. This bi-directional interplay between CAFs and tumor cells within the tumor microenvironment ends up fostering tumor cancer progression. CAFs are therefore key regulators of tumor progression and represent an under-explored therapeutic target in thyroid cancer.Fil: Fozzatti, Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Cheng Sheue-yann. National Institutes of Health; Estados Unido

The effect of the global postural reeducation (GPR) in female stress urinary incintinence

Orientadores: Paulo Cesar Rodrigues Palma, Miriam DambrosDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências MédicasResumo: A Incontinência urinária de esforço (IUE) feminina, condição com alta prevalência, é definida como um sinal e sintoma ligado a distúrbios funcionais da uretra (esfíncteres) e/ou bexiga e não caracteriza uma doença. Trata-se então de uma disfunção mecânica em que alterações na biomecânica da bacia pélvica podem estar associadas à modificação deste mecanismo. Atualmente tem-se valorizado e vem-se aplicando o tratamento fisioterapêutico nesta afecção, como o treinamento dos músculos do assoalho pélvico, obtendo-se bons resultados a curto e médio prazos. Além disso, técnicas baseadas na abordagem global da paciente, que consideram aspectos da estrutura postural, estão ainda em fase de investigação. O trabalho aqui descrito constou da aplicação do tratamento da Reeducação Postural Global (RPG), trabalhando-se a reestruturação postural por meio do reequilíbrio do Sistema músculo esquelético (SME), alongamento das cadeias musculares e reequilíbrio dos eixos ósseos, num enfoque global. Objetivo: Avaliar os efeitos da RPG nas queixas de IUE e qualidade de vida em um grupo de mulheres incontinentes. Casuística e Método: Para o estudo, foram selecionadas 26 mulheres portadoras de queixa clínica de IUE, que foram submetidas ao tratamento da RPG. O tratamento constou de sessões semanais de 50 minutos num período de três meses e posteriormente de sessões quinzenais por mais três meses. O grupo foi acompanhado por seis meses após final do tratamento, sendo reavaliado no término do tratamento, no terceiro e sexto meses. A avaliação foi feita usando Questionário de Qualidade de Vida, diário miccional de três dias, Pad Use e Avaliação funcional do assoalho pélvico (AFA). No término do tratamento e no seguimento de seis meses, as pacientes também foram avaliadas por meio de escala analógica de satisfação. Resultados: Das 26 pacientes que iniciaram o programa, 25 concluíram o tratamento. No final deste quatro pacientes (16%) estavam curadas, 18 (72%) apresentaram melhora significativa e três (12%) não apresentaram melhora. No seguimento de seis meses, seis (24%) pacientes estavam curadas, 16 (64%) apresentaram melhora e três (12%) não apresentaram melhora. Diferenças significativas foram notadas no número de perdas (p<0.001), Pad Use (p<0.001) e AFA (p<0.001). Além disso, foi percebida melhora em todos os domínios do Questionário de Qualidade de Vida, especialmente em Percepção geral da saúde (p<0.005) e Impacto da incontinência (p<0.001) em todos os seguimentos da avaliação. Conclusão: A RPG induziu à melhora significativa dos sintomas de IUE e qualidade de vida no grupo de mulheres incontinentes estudadoAbstract: Stress Urinary Incontinence (SUI), is defined as a signal and/or a symptom connected to functional disorders of the urethra (sphincter) and/or blader and do not characterize a disease. It is, indeed a dysfunction where alterations in the pelvis biomechanics can be associated to a modification of this mechanism. Lately, physiotherapeutic treatments, as the training of the pelvic floor muscles, have been used and good results have been obtained in short and medium terms. Besides, techniques based on a global approach of the patient, which consider also aspects of the postural structure, are still under investigation. The work hereby described consisted of the application of the Global Postural Reeducation (GPR) treatment, in which the postural restructuring is worked through the reestablishment of the balance of the skeletal muscle system, stretching of the muscle chains and rebalance of the bone axis, in a global approach. Objective: Evaluate the effects of the GPR on Stress Urinary Incontinence and Life Quality in a group of incontinent women. Material and Methods: For this study, 26 women with SUI were selected, who underwent a GPR treatment. All patients were treated for six months using GPR, 50 minutes weekly sessions during three months and three more months of sessions every other week. The patients were evaluated before and after the treatment, and also at three and six months follow-up. Outcome measures were made using King's Health Questionnaire, three days voiding diary, Pad Use and Functional Evaluation of Pelvic Floor (FEPF). In the end of the treatment and after six months, the patient satisfaction was evaluated trough a standardized analogical visual scale. Results: Twenty-five patients were available for follow-up. At the end of the treatment there were four (16%) patients completely dry, 18 (72%) pesented significant improvement and three (12%) did not presented improvement. At six months follow-up there were six (24%) patients completely dry, 16 (64%) improved and three (12%) failures. Significant differences were noted in the number of leak episodes (p<0.001), Pad Use (p<0.001) and FEPF (p<0.001). Regarding the King's Health Questionnaire, improvement in all domains were observed, especially in General Perception of Health (p<0.005), leakage impact (p<0.001) in all the moments of evaluation. Conclusion: GPR significantly improved the symptoms and Quality of Life in women with SUIMestradoPesquisa ExperimentalMestre em Cirurgi

Female urinary incontinence treatment using global postural re-education (GPR) : longitudinal comparative study

Orientadores: Viviane Herrmann, Paulo César Rodrigues PalmaTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências MédicasResumo: A incontinência urinária de esforço é uma das queixas clínicas mais comuns de mulheres em consultório médico e o desequilíbrio postural tem sido considerado como possível causa deste sintoma. Estudos demonstram que disfunções posturais como a hiperlordose, a anteversão da bacia pélvica, dores lombares e fraqueza dos músculos abdominais estão associados às disfunções dos músculos do assoalho pélvico. Objetivo: Comparar o efeito da Reeducação Postural Global com o treinamento dos músculos do assoalho pélvico no tratamento da incontinência urinária de esforço feminina a curto e a longo prazo. Metodologia: Cinquenta e duas mulheres com queixa clínica de incontinência urinária de esforço foram divididas em dois grupos: o Grupo 1 (G1) recebeu tratamento pela Reeducação Postural Global em 1 sessão semanal de 50 minutos por três meses e o Grupo 2 (G2) recebeu treinamento dos músculos do assoalho pélvico quatro vezes por semana, uma vez supervisionada em sessão individual de 50 minutos e três não supervisionadas, por três meses. Ao final do tratamento, após 6 meses, e em dois anos, foi realizada avaliação subjetiva (cura, melhora, inalterada e piora), diário miccional, avaliação funcional do assoalho pélvico (AFA) e questionário de qualidade de vida. Resultados: Concluíram o tratamento 25 pacientes do G1 e 17 do G2. A avaliação subjetiva do G1 ao final do tratamento apresentou 16% cura e após seis meses, 24%. No G2, no final do tratamento, nenhuma paciente considerava-se curada, 69% referiram melhora e 31% encontravam-se inalteradas. Após seis meses, 19% referiram cura, 37,5% melhoram, 31% inalteradas e 12,5% que haviam referido melhora ao final do tratamento, apresentaram piora dos sintomas. O número de episódios de perda e o número de troca de absorventes diminuiram significativamente nos dois grupos (p<0,001), sendo significativamente menor no G1. A Avaliação Funcional do Assoalho Pélvico (AFA) melhorou significativamente nos dois grupos (p<0,001), sem diferença entre eles. A avaliação da Qualidade de Vida demonstrou melhora significativa nos dois grupos, em todos os domínios. Após dois anos foram reavaliadas 21 pacientes no G1 e 12 no G2. O G1 apresentou 47,6% de cura e o G2, 16,7%, sendo que neste grupo, 33,3% das pacientes referiram piora com relação ao final do tratamento. O número de perdas e o número de absorventes diminuíram significativamente nos dois grupos (p=0,0001), sem diferença significativa entre eles (p=0,0787 e p=0,0579, respectivamente). A AFA melhorou significativamente no G1, porém no G2, apresentou melhora significativa ao final do tratamento e mante-se inalterada no seguimento de dois anos (p=0,045). Conclusão: A RPG mostrou-se uma alternativa eficaz no tratamento da Incontinência Urinária de Esforço Feminina, com resultados comparáveis ao treinamento dos músculos do assoalho pélvico, a curto e a longo prazoAbstract: Stress urinary incontinence (SUI) is one of the most common complains of women and postural unbalances have been considered as a possible cause. Studies have shown that postural disequilibrium such as hiperlordose, pelvis anteversion, lumbar pain and weakness of the abdominal muscles are associated to pelvic floor muscles dysfunctions. Objective: To compare the effect of Global Postural Re-education (GPR) and Pelvic Floor Muscle Training (PFMP) for the treatment of female stress urinary incontinence at short and long term. Methodology: Fifty-two women with SUI complain were distributed into two groups: Group 1 (G1) was submitted to 50 minutes weekly sessions of GPR for three months and Group 2 (G2) performed PFMT four times a week for three months, being one time in individual session under professional supervision and the other three times at home, for three months. Patients were evaluated at the end of the treatment and after six months and two years on subjective evaluation (cure, improvement, no change and recurrence), voiding diary, functional evaluation of pelvic floor (FEPF) and through a questionnaire of quality of life. Results: The number of patients completing the treatments was 25 in G1 and 17 in G2. The subjective evaluation of G1 showed cure in 16% women at the end of the treatment and 24% after six months. In G2 no women reported cure at the end of the treatment, 69% indicated improvement and 31% reported no change. After six months, the reports of cure, improvement and no change in G2 were, respectively, 19%, 37.5% and 31%. Additionally, 12.5% of the women in this group, who had referred to improvement at the end of the treatment, reported symptom recurrence after six months. The urine leakage episodes reduced significantly in both groups (p<0.001), being significantly lower in G1. PAD use reduced significantly in both groups (p<0.001) and was significantly lower in G1. FEPF improved significantly in both groups (p<0.001), with no difference between them. The evaluation of Quality of Life had shown significantly improvement in both groups, in all domains. Two years after the end of the treatment, 21 patients of G1and 12 of G2 were reevaluated. The amount of women reporting cure in G1 and G2 were, respectively, 47.6% and 16.7%. In G2, 33.3% of the patients referred to symptoms recurrence in comparison with the end of the treatment. Episodes of urine leakage and PAD use reduced significantly in both groups (p=0.0001), without significant difference between groups (p=0.0787 and p=0.0579, respectively). FEPF had shown improvement in G1 in the evaluation at six months and two years; however, in G2 it had shown improvement at the end of the treatment but it did not change in the evaluation at two years. This shows that the behavior of the groups was different throughout the time (p=0.045). Conclusion: GPR has proven to be an efficient alternative to treat SUI in women when compared to PFMT, either on short term or long term follow-upDoutoradoFisiopatologia CirúrgicaDoutor em Ciência

Oncogenic Actions of the Nuclear Receptor Corepressor (NCOR1) in a Mouse Model of Thyroid Cancer

Studies have suggested that the nuclear receptor corepressor 1 (NCOR1) could play an important role in human cancers. However, the detailed molecular mechanisms by which it functions in vivo to affect cancer progression are not clear. The present study elucidated the in vivo actions of NCOR1 in carcinogenesis using a mouse model (ThrbPV/PV mice) that spontaneously develops thyroid cancer. ThrbPV/PV mice harbor a dominantly negative thyroid hormone receptor b (TRb) mutant (denoted as PV). We adopted the loss-of-the function approach by crossing ThrbPV mice with mice that globally express an NCOR1 mutant protein (NCOR1DID) in which the receptor interaction domains have been modified so that it cannot interact with the TRb, or PV, in mice. Remarkably, expression of NCOR1DID protein reduced thyroid tumor growth, markedly delayed tumor progression, and prolonged survival of ThrbPV/PVNcor1DID/DID mice. Tumor cell proliferation was inhibited by increased expression of cyclin-dependent kinase inhibitor 1 (p21waf1/cip1; Cdkn1A), and apoptosis was activated by elevated expression of pro-apoptotic BCL-Associated X (Bax). Further analyses showed that p53 was recruited to the p53- binding site on the proximal promoter of the Cdkn1A and the Bax gene as a co-repressor complex with PV/NCOR1/histone deacetylas-3 (HDAC-3), leading to repression of the Cdkn1A as well as the Bax gene in thyroids of ThrbPV/PV mice. In thyroids of ThrbPV/PVNcor1DID/DID mice, the p53/PV complex could not recruit NCOR1DID and HDAC-3, leading to de-repression of both genes to inhibit cancer progression. The present studies provided direct evidence in vivo that NCOR1 could function as an oncogene via transcription regulation in a mouse model of thyroid cancer.Fil: Fozzatti, Laura. Consejo Nacional de Invest.cientif.y Tecnicas. Centro Cientifico Tecnol.conicet - Cordoba. Instituto de Inv. Medicas Mercedes y Martin Ferreyra;Fil: Park, Jeong Won.Fil: Li, Zhao.Fil: Willingham, Mark C..Fil: Cheng, Sheue-yann

Trypanosoma cruzi Exploits Wnt Signaling Pathway to Promote Their Intracellular Replication in Macrophages

During the acute phase of Trypanosoma cruzi infection, macrophages can act as host cells for the parasites as well as effector cells in the early anti-parasitic immune response. Thus, the targeting of specific signaling pathways could modulate macrophages response to restrict parasite replication and instruct an appropriate adaptive response. Recently, it has become evident that Wnt signaling has immunomodulatory functions during inflammation and infection. Here, we tested the hypothesis that during T. cruzi infection, the activation of Wnt signaling pathway in macrophages plays a role in modulating the inflammatory/tolerogenic response and therefore regulating the control of parasite replication. In this report, we show that early after T. cruzi infection of bone marrow-derived macrophages (BMM), β-catenin was activated and Wnt3a, Wnt5a, and some Frizzled receptors as well as Wnt/β-catenin pathway's target genes were upregulated, with Wnt proteins signaling sustaining the activation of Wnt/β-catenin pathway and then activating the Wnt/Ca+2 pathway. Wnt signaling pathway activation was critical to sustain the parasite's replication in BMM; since the treatments with specific inhibitors of β-catenin transcriptional activation or Wnt proteins secretion limited the parasite replication. Mechanistically, inhibition of Wnt signaling pathway armed BMM to fight against T. cruzi by inducing the production of pro-inflammatory cytokines and indoleamine 2,3-dioxygenase activity and by downregulating arginase activity. Likewise, in vivo pharmacological inhibition of the Wnts' interaction with its receptors controlled the parasite replication and improved the survival of lethally infected mice. It is well established that T. cruzi infection activates a plethora of signaling pathways that ultimately regulate immune mediators to determine the modulation of a defined set of effector functions in macrophages. In this study, we have revealed a new signaling pathway that is activated by the interaction between protozoan parasites and host innate immunity, establishing a new conceptual framework for the development of new therapies.Fil: Volpini, Ximena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Ambrosio, Laura Fernanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Fozzatti, Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Insfran, Constanza. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Stempin, Cinthia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Cervi, Laura Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Motran, Claudia Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentin

Interplay of fibroblasts with anaplastic tumor cells promotes follicular thyroid cancer progression

Thyroid cancer is the most common endocrine malignancy. Anaplastic thyroid cancer is one of the most aggressive thyroid tumors. It is known that activation of oncogenes and/or inactivation of tumor suppressor genes in tumor cells promotes tumorigenesis. The microenvironment of the tumor also plays a key role on cancer development and progression in a variety of tumors. However, the mechanisms by which tumor-stroma crosstalk in thyroid cancer remains poorly characterized. In this study we aimed to understand how interactions between fibroblasts and anaplastic thyroid cancer cells contribute to thyroid carcinogenesis. We first characterized the phenotypic changes of human fibroblasts in vitro through co-cultures by using transwells as well as by using anaplastic thyroid cancer cells-derived conditioned media. We found that fibroblasts acquired an activated phenotype or also known as cancer-associated fibroblast phenotype after being in contact with soluble factors secreted from anaplastic thyroid cancer cells, compared to the fibroblasts in mono-cultures. All the changes were partly mediated through Src/Akt activation. Treatment with the antioxidant N-acetyl-cysteine reversed in part the metabolic phenotype of activated fibroblasts. Remarkably, conditioned media obtained from these activated fibroblasts promoted cell proliferation and invasion of follicular thyroid cancer cell line, FTC-133 cells. Thus, a reciprocal and dynamic interaction exists between tumor and stromal cells, which results in the promotion of thyroid tumorigenesis. The present studies have advanced the understanding of the molecular basis of tumor-stroma communications, enabling identification and targeting of tumor-supportive mechanisms for novel treatment modalities.Fil: Fozzatti, Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Alamino, Vanina Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Park, Sunmi. National Institutes of Health; Estados UnidosFil: Giusiano, Lucila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Volpini, Ximena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Zhao, Li. National Institutes of Health; Estados UnidosFil: Stempin, Cinthia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Donadio, Ana Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Cheng, Sheue-yann. National Institutes of Health; Estados UnidosFil: Pellizas, Claudia Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentin

Secreted factors by Anaplastic Thyroid Cancer Cells Induce Tumor-Promoting M2-like Macrophage Polarization through a TIM3-Dependent Mechanism

Anaplastic thyroid cancer (ATC) is a highly aggressive type of thyroid cancer (TC). Currently, no effective target treatments are available that can improve overall survival, with ATC representing a major clinical challenge because of its remarkable lethality. Tumor-associated macrophages (TAMs) are the most evident cells in ATCs, and their high density is correlated with a poor prognosis. However, the mechanisms of how TAMs promote ATC progression remain poorly characterized. Here, we demonstrated that the treatment of human monocytes (THP-1 cells) with ATC cell-derived conditioned media (CM) promoted macrophage polarization, showing high levels of M2 markers. Furthermore, we found that STAT3 was activated, and this was correlated with an increased expression and secretion of the inflammatory cytokine interleukin-6. Remarkably, the M2-like macrophages obtained revealed tumor-promoting activity. A cytokine array analysis demonstrated that M2-like macrophage-derived CM contained high levels of TIM3, which is an important immune regulatory molecule. Consistently, TIM3 expression was up-regulated in THP-1 cells cultured with ATC cell-derived CM. Moreover, TIM3 blockade significantly reversed the polarization of THP-1 cells induced by ATC cell-secreted soluble factors. We validated the clinical significance of the TIM3 in human TC by analyzing public datasets and found that the expression of TIM3 and its ligand galectin 9 was significantly higher in human TC tissue samples than in normal thyroid tissues. Taken together, our findings identified a new mechanism by which TIM3 induces tumor-promoting M2-like macrophage polarization in TC. Furthermore, TIM3 interference might be a potential tool for treatment of patients with ATC.Fil: Stempin, Cinthia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; ArgentinaFil: Geysels, Romina Celeste. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; ArgentinaFil: Park, Sunmi. National Institutes of Health; Estados UnidosFil: Palacios, Luz María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; ArgentinaFil: Volpini, Ximena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; ArgentinaFil: Motran, Claudia Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; ArgentinaFil: Acosta Rodriguez, Eva Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; ArgentinaFil: Nicola, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; ArgentinaFil: Cheng, Sheue Yann. National Institutes of Health; Estados UnidosFil: Pellizas, Claudia Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; ArgentinaFil: Fozzatti, Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; Argentin

COVID-19 patients display changes in lymphocyte subsets with a higher frequency of dysfunctional CD8lo T cells associated with disease severity

This work examines cellular immunity against SARS-CoV-2 in patients from Córdoba, Argentina, during two major waves characterized by different circulating viral variants and different social behavior. Using flow cytometry, we evaluated the main lymphocyte populations of peripheral blood from hospitalized patients with moderate and severe COVID-19 disease. Our results show disturbances in the cellular immune compartment, as previously reported in different cohorts worldwide. We observed an increased frequency of B cells and a significant decrease in the frequency of CD3+ T cells in COVID-19 patients compared to healthy donors (HD). We also found a reduction in Tregs, which was more pronounced in severe patients. During the first wave, the frequency of GZMB, CD107a, CD39, and PD-1-expressing conventional CD4+ T (T conv) cells was significantly higher in moderate and severe patients than in HD. During the second wave, only the GZMB+ T conv cells of moderate and severe patients increased significantly. In addition, these patients showed a decreased frequency in IL-2-producing T conv cells. Interestingly, we identified two subsets of circulating CD8+ T cells with low and high CD8 surface expression in both HD and COVID-19 patients. While the percentages of CD8hi and CD8lo T cells within the CD8+ population in HD are similar, a significant increase was observed in CD8lo T cell frequency in COVID-19 patients. CD8lo T cell populations from HD as well as from SARS-CoV-2 infected patients exhibited lower frequencies of the effector cytokine-producing cells, TNF, IL-2, and IFN-γ, than CD8hi T cells. Interestingly, the frequency of CD8lo T cells increased with disease severity, suggesting that this parameter could be a potential marker for disease progression. Indeed, the CD8hi/CD8lo index helped to significantly improve the patient’s clinical stratification and disease outcome prediction. Our data support the addition of, at least, a CD8hi/CD8lo index into the panel of biomarkers commonly used in clinical labs, since its determination may be a useful tool with impact on the therapeutic management of the patients