Ovarian Cancer-Associated Ascites Have High Proportions of Cytokine-Responsive CD56bright NK Cells.

Ovarian cancer is the most lethal gynecological malignancy, with serous histotype as the most prevalent epithelial ovarian cancer (EOC). Peritoneal ascites is a frequent comorbidity in advanced EOC. EOC-associated ascites provide a reliable sampling source for studying lymphocytes directly from tumor environment. Herein, we carried out flow cytometry-based analysis to readdress issues on NK and T lymphocyte subsets in women with advanced EOC, additionally evaluating phenotypic modulation of their intracellular pathways involved in interleukin (IL)-2 and IL-15 signaling. Results depicted ascites as an inflammatory and immunosuppressive environment, presenting significantly (p < 0.0001) higher amounts of IL-6 and IL-10 than in the patients' blood, as well as significantly (p < 0.05) increased expression of checkpoint inhibitory receptors (programmed death protein-1, PD-1) and ectonucleotidase (CD39) on T lymphocytes. However, NK lymphocytes from EOC-associated ascites showed higher (p < 0.05) pS6 phosphorylation compared with NK from blood. Additionally, in vitro treatment of lymphocytes with IL-2 or IL-15 elicited significantly (p < 0.001) phosphorylation of the STAT5 protein in NK, CD3 and CD8 lymphocytes, both from blood and ascites. EOC-associated ascites had a significantly (p < 0.0001) higher proportion of NK CD56bright lymphocytes than blood, which, in addition, were more responsive (p < 0.05) to stimulation by IL-2 than CD56dim NK. EOC-associated ascites allow studies on lymphocyte phenotype modulation in the tumor environment, where inflammatory profile contrasts with the presence of immunosuppressive elements and development of cellular self-regulating mechanisms.

Preservation of cytotoxic granule production in response to mycobacterial antigens by T-lymphocytes from vertically HIV-infected Brazilian youth on effective combined antiretroviral therapy

ABSTRACT Background: HIV infection harms adaptive cellular immunity mechanisms. Long-term virological control by combined antiretroviral therapy (cART) reduces the risk of mycobacterial infections. Thus, we aimed to study cellular responses to mycobacterial antigens in 20 HIV-infected adolescents with at least one year of virological control (HIV-RNA <40 copies/mL) and 20 healthy adolescents. Methods: We evaluated CD8 and γδ T-cell degranulation by measurement of CD107a membrane expression after stimulation with lysates from BCG (10 µg/mL) and H37RA Mycobacterium tuberculosis (Mtb, 10 µg/mL). Immune activation and antigen-presenting ability were also assessed by determination of HLA-DR, CD80, and CD86 markers. Results: TCR γδ T-cell CD107a expression was similar between groups in response to mycobacterial antigens, and lower in the HIV-infected group in response to mitogen. Higher baseline HLA-DR expression and lower mycobacterial-stimulated expression was found within the HIV-infected group. Conclusions: Similar degranulation in stimulated CD8+ and TCR γδ T-cells from HIV-infected adolescents, when compared to healthy controls suggests long-term immunological preservation with immune reconstitution under successful cART. However, differences in HLA-DR expression may represent ongoing inflammation and lower specific responses in HIV-infected youth. These features may be relevant in the context of the precocity and severity of vertically acquired HIV infection.

Terapia celular combinada com membranas de biopolímeros melhora a cicatrização de úlceras em paciente com dermatomiosite juvenil / Combining cell therapy with biopolymer films improves wound healing in a juvenile dermatomyositis patient

Introdução: Dermatomiosite juvenil (DMJ) é doença sistêmica que afeta a musculatura proximal e a pele de crianças. A doença ulcerada é um desafio terapêutico. Objetivo: Avaliar a melhora da doença ulcerada na DMJ, pelo uso de terapia celular. Métodos: Realização de cocultura de fibroblastos e queratinócitos autólogos e aplicação dessas células nas úlceras juntamente com cola de fibrina e colocação de membrana de quitosana-alginato ou quitosana-xantana sobre as lesões. Resultados: Menos de 12 horas após a terapia, o paciente referiu completa eliminação da dor e, dentro de dois dias, estava presente tecido de cicatrização. Algumas das úlceras estavam quase completamente cicatrizadas no final da primeira semana, e algumas das calcinoses desapareceram. Essa técnica não cura a doença, mas melhora a qualidade de vida, sendo possível criopreservar as células saudáveis do paciente para tratar novas lesões. Sendo as células de origem autóloga, elimina-se o risco de rejeição. Além disso, esse procedimento não necessita de debridamento das lesões nem hospitalização. Conclusões: A aplicação de culturas autólogas de fibroblastos e queratinócitos em úlceras já é considerada tratamento efetivo em pacientes com queimaduras e outras feridas cutâneas e, agora mostrou-se também eficaz no tratamento de feridas na DMJ.

Introduction: Juvenile dermatomyositis (JDM) is a systemic disease that affects children's proximal musculature and skin. The ulcerated stage of the disease is a therapeutic challenge. Objective: To evaluate the improvement of ulcerated stage of JDM caused by the use of cell therapy. Methods: Co-culture of autologous fibroblasts and keratinocytes, application of these cells in ulcers in conjunction with fibrin glue, and placement of chitosan-alginate or chitosan-xanthan membrane on the lesions. Results: Less than 12 hours after therapy, the patient reported complete cessation of pain and, within 2 days, healing tissue emerged. Some of the ulcers were almost completely healed by the end of the 1st week, and some of the calcinoses disappeared. This technique does not cure the disease, however it improves the patient's quality of life, and it is possible to cryopreserve healthy cells to treat new lesions. Given the fact that the cells are of autologous origin, the risk of rejection is eliminated. Furthermore, this procedure does not require debridement of the lesions or hospitalization. Conclusions: The application of autologous cultures of fibroblasts and keratinocytes in ulcers is already considered an effective treatment in patients with burns and other skin wounds, and has now also been proven effective in the treatment of wounds in JDM.

Platelet-rich plasma (PRP) and adipose-derived mesenchymal stem cells: stimulatory effects on proliferation and migration of fibroblasts and keratinocytes in vitro.

The clinical use of tissue engineering associated with cell therapy is considered a new alternative therapy for the repair of chronic lesions with potential application in different medical areas, mostly in orthopedic and dermatological diseases. Platelet-rich plasma (PRP) is a rich source of growth factors and cytokines important for wound healing. Adipose-derived mesenchymal stem cells (ADSCs) have shown potential to accelerate the resolution of ulcers, to stimulate cell proliferation, and to benefit the quality of skin repair. This study aims to determine the effect of PRP and conditioned medium (CM) from ADSC on fibroblast and keratinocyte proliferation in vitro. Migration and proliferation assays were performed to evaluate the growth of fibroblasts and keratinocytes in the presence of PRP, CM, and CM + PRP. Significant proliferative stimulation was observed after 48 h of culture (p < 0.05) on mean absorbance of fibroblasts cultured with 10 and 25 % PRP, 100 % CM, and 25 % PRP + 25 % CM, if compared with control. Keratinocyte proliferation was stimulated after 48 h in cultures with 25, 50, and 100 % CM, and growth was compared with controls. The migration assay detected a significant migratory stimulus in fibroblasts cultured with 10 % PRP + 10 % CM after 48 h. These in vitro results suggest that PRP and ADSC have therapeutic potential for healing and re-epithelialization of chronic wounds in vivo.