Leishmania (Viannia) braziliensis amastigotes induces the expression of TNFα and IL-10 by human peripheral blood mononuclear cells in vitro in a TLR4-dependent manner.

While the role of Toll-like receptors (TLRs) has been investigated in murine models of tegumentary leishmaniasis caused by Leishmania (Viannia) braziliensis, the interaction between TLRs and Leishmania sp. has not been investigated in human cells. The aim of this study was to evaluate the involvement of TLR4 in cytokine production of human peripheral blood mononuclear cells (PBMCs) induced by L. braziliensis, and whether the parasite alters the expression of TLR4 on monocytes/macrophages. Amastigote forms were obtained from mice lesions and PBMCs were isolated from healthy donors. PBMCs were cultured in absence or presence of IFNγ, TLR4 neutralizing antibodies, natural antagonist of TLR4 (Bartonella LPS), TLR4 agonist (E. coli LPS), and amastigote forms. The concentrations of tumor necrosis factor (TNFα) and interleukin 10 (IL-10) were assayed by ELISA and TLR4 expression by flow cytometry. Amastigotes forms of L. braziliensis induced TNFα and IL-10 production only in IFNγ-primed PBMCs. The TNFα and IL-10 production was inhibited by TLR4 neutralization, both with anti-TLR4 antibodies and Bartonella LPS. Interestingly, addition of E. coli LPS further increased TNFα but not IL-10 production induced by L. braziliensis amastigotes. Amastigotes of L. braziliensis strongly reduced membrane TLR4 expression on monocytes/macrophages, apparently by internalization after the infection. The present study reveals that TLR4 drives the production of TNFα and IL-10 induced by L. braziliensis amastigotes and that the parasites decrease TLR4 expression on monocyte surface.

Interleukin 32γ (IL-32γ) is highly expressed in cutaneous and mucosal lesions of American Tegumentary Leishmaniasis patients: association with tumor necrosis factor (TNF) and IL-10.

BACKGROUND: The interleukin 32 (IL-32) is a proinflammatory cytokine produced by immune and non-immune cells. It can be induced during bacterial and viral infections, but its production was never investigated in protozoan infections. American Tegumentary Leishmaniasis (ATL) is caused by Leishmania protozoan leading to cutaneous, nasal or oral lesions. The aim of this study was to evaluate the expression of IL-32 in cutaneous and mucosal lesions as well as in peripheral blood mononuclear cells (PBMC) exposed to Leishmania (Viannia) braziliensis. METHODS: IL-32, tumour necrosis factor (TNF) and IL-10 protein expression was evaluated by immunohistochemistry in cutaneous, mucosal lesions and compared to healthy specimens. The isoforms of IL-32α, ß, δ, γ mRNA, TNF mRNA and IL-10 mRNA were assessed by qPCR in tissue biopsies of lesions and healthy skin and mucosa. In addition, PBMC from healthy donors were cultured with amastigotes of L. (V.) braziliensis. In lesions, the parasite subgenus was identified by PCR-RFLP. RESULTS: We showed that the mRNA expression of IL-32, in particular IL-32γ was similarly up-regulated in lesions of cutaneous (CL) or mucosal (ML) leishmaniasis patients. IL-32 protein was produced by epithelial, endothelial, mononuclear cells and giant cells. The IL-32 protein expression was associated with TNF in ML but not in CL. IL-32 was not associated with IL-10 in both CL and ML. Expression of TNF mRNA was higher in ML than in CL lesions, however levels of IL-10 mRNA were similar in both clinical forms. In all lesions in which the parasite was detected, L. (Viannia) subgenus was identified. Interestingly, L. (V.) braziliensis induced only IL-32γ mRNA expression in PBMC from healthy individuals. CONCLUSIONS: These data suggest that IL-32 plays a major role in the inflammatory process caused by L. (Viannia) sp or that IL-32 is crucial for controlling the L. (Viannia) sp infection.

Prevalence and trends in transfusion-transmissible infections among blood donors in Brazil from 2010 to 2016.

BACKGROUND: Assessing trends in the rate of transfusion-transmissible infections (TTIs) in blood donors is critical to the monitoring of the blood supply safety and the donor screening effectiveness. The objective of this study was to conduct a trend analysis of TTIs and associated demographic factors of donors at a public blood bank in the central Brazil. METHODS: A retrospective analysis (2010-2016) of blood donation data was performed to determine the prevalence of markers for TTIs. Multinomial and multivariate logistic regression were used to verify the association between the explanatory variables and TTIs. The trend was evaluated with the Prais Winsten's regression analysis. RESULTS: The prevalence of TTIs was 4.04% (5,553 donors) among 137,209 donors, with a steady trend in the analyzed period. The seroprevalence for the hepatitis B virus (HBV), syphilis, hepatitis C virus (HCV), human immunodeficiency virus (HIV), Chagas disease, and human T-lymphotropic virus (HTLV) were 1.63%, 0.87%, 0.46%, 0.21%, 0.21% and 0.09%, respectively. The prevalence of HBV decreased (b = -0.021, p <  0.001), while syphilis increased (b = 0.112; p =  0.001), during the period investigated. The probability for a positive test for TTI was higher among donors with a low level of education, aged ≥30 years old, without stable marital status, and first-time donors. CONCLUSIONS: Even with the reduction in HBV, the increased rate of syphilis may have contributed to the fact that the overall rate of TTIs did not decrease in the evaluated period.

Red blood cell alloimmunization among hospitalized patients: transfusion reactions and low alloantibody identification rate.

BACKGROUND: Unexpected red blood cell alloantibodies can cause hemolytic transfusion reactions. In this study, the prevalence of alloimmunization, the rate of identification of alloantibodies and the rate of blood transfusion reactions among transfused patients were identified in a clinical emergency hospital in Brazil. METHODS: Transfusions and clinical records of patients who had a positive indirect antiglobulin test between January and December 2013 were analyzed. RESULTS: Of 1169 patients who received blood transfusions, 28 had positive indirect antiglobulin tests, with one patient having two positive tests at different times, resulting in 29 positive tests during the period of this study. Alloantibodies were identified in 58.6% (17/29) of the cases. In 27.5% (8/29), identification was inconclusive and it was not possible to confirm alloimmunization. The rate of red blood cell alloimmunization was 1.71% (21/1169). Of 21 cases of alloimmunization, four (19%) were unidentified due to an unusual agglutination profile. All identified alloantibodies were clinically significant (10/17 anti-Rh, 5/17 anti-Kell and 2/17 anti-MNS). In two patients who had positive indirect antiglobulin tests, one had an unidentified alloantibody, and the other had an inconclusive test and developed a hemolytic transfusion reaction. CONCLUSION: The prevalence of clinically important red blood cell alloantibodies and hemolytic transfusion reactions among patients with unidentified alloantibodies suggests that specific laboratory techniques should be performed to identify alloantibodies in cases of pan-reactivity or autoantibodies to improve transfusion safety.

Redução de doações de sangue: a importância da doação de campanha em Goiânia, Estado de Goiás, Brasil / Reduction in blood donations: the importance of campaing donation in Goiânia, Goiás, Brazil

Analisar as tendências das doações de sangue no Hemocentro do Estado de Goiás (HEMOGO), Brasil, considerando as campanhas de incentivo e as infecções transmissíveis por transfusão. Estudo retrospectivo das doações de sangue entre 2010-2016. Os doadores foram agrupados em categorias autóloga, voluntária, de reposição e de campanha. Houve 149.983 doações com redução de 29% (p <0,05). As doações por homens, com idade entre 18 e 29 anos e com menor escolaridade diminuíram (p <0,05) ao longo do tempo. Quase 50% das doações eram da categoria voluntária, 30% de campanha, 18% de reposição e 1% de outras categorias. As doações da campanha diminuíram 5,02% (p <0,05) entre 2010 a 2016. A prevalência de infecções transmitidas por transfusão (ITT) foi de 3,71% e a chance de doadores de campanha terem ITT foi menor (OR = 0,8628; IC: 0,8126 - 0,9161; p <0,0001). Os resultados mostraram uma redução nas doações de sangue, influenciadas principalmente por uma diminuição nas doações da campanha.

To analyze the trends of blood donations in a public blood center of Goias, Brazil (HEMOGO ­ Hemocentro do Estado de Goiás), considering incentive campaigns and transfusion transmissible infection. Retrospective study of the blood donations between 2010-2016. Donors were grouped into autologous, voluntary, replacement and campaign categories. There were 149,983 donations with a reduction of 29% (p<0.05) in the investigated period. Donations by males, aging between 18 and 29 years old, and those with a lower level of education decreased (p<0.05) over time. Almost 50% of donations were from the voluntary category, 30% from campaing, 18% from replacement and 1% from other categories. The campaign donations decreased 5.02% (p<0.05) during the 2010 to 2016. The prevalence of transfusion-transmitted infections (TTI) was 3.71% and the chance of campaign donors having TTI was lower (OR = 0.8628; CI: 0.8126 - 0.9161; p<0.0001). The results showed a significant reduction in the number of blood donations mainly influenced by a decrease in campaign donations.

Prevalence and trends in transfusion-transmissible infections among blood donors in Brazil from 2010 to 2016

ABSTRACT Background: Assessing trends in the rate of transfusion-transmissible infections (TTIs) in blood donors is critical to the monitoring of the blood supply safety and the donor screening effectiveness. The objective of this study was to conduct a trend analysis of TTIs and associated demographic factors of donors at a public blood bank in the central Brazil. Methods: A retrospective analysis (2010-2016) of blood donation data was performed to determine the prevalence of markers for TTIs. Multinomial and multivariate logistic regression were used to verify the association between the explanatory variables and TTIs. The trend was evaluated with the Prais Winsten's regression analysis. Results: The prevalence of TTIs was 4.04% (5,553 donors) among 137,209 donors, with a steady trend in the analyzed period. The seroprevalence for the hepatitis B virus (HBV), syphilis, hepatitis C virus (HCV), human immunodeficiency virus (HIV), Chagas disease, and human T-lymphotropic virus (HTLV) were 1.63%, 0.87%, 0.46%, 0.21%, 0.21% and 0.09%, respectively. The prevalence of HBV decreased (b = −0.021, p < 0.001), while syphilis increased (b = 0.112; p = 0.001), during the period investigated. The probability for a positive test for TTI was higher among donors with a low level of education, aged ≥30 years old, without stable marital status, and first-time donors. Conclusions: Even with the reduction in HBV, the increased rate of syphilis may have contributed to the fact that the overall rate of TTIs did not decrease in the evaluated period.

Red blood cell alloimmunization among hospitalized patients: transfusion reactions and low alloantibody identification rate

ABSTRACT Background: Unexpected red blood cell alloantibodies can cause hemolytic transfusion reactions. In this study, the prevalence of alloimmunization, the rate of identification of alloantibodies and the rate of blood transfusion reactions among transfused patients were identified in a clinical emergency hospital in Brazil. Methods: Transfusions and clinical records of patients who had a positive indirect antiglobulin test between January and December 2013 were analyzed. Results: Of 1169 patients who received blood transfusions, 28 had positive indirect antiglobulin tests, with one patient having two positive tests at different times, resulting in 29 positive tests during the period of this study. Alloantibodies were identified in 58.6% (17/29) of the cases. In 27.5% (8/29), identification was inconclusive and it was not possible to confirm alloimmunization. The rate of red blood cell alloimmunization was 1.71% (21/1169). Of 21 cases of alloimmunization, four (19%) were unidentified due to an unusual agglutination profile. All identified alloantibodies were clinically significant (10/17 anti-Rh, 5/17 anti-Kell and 2/17 anti-MNS). In two patients who had positive indirect antiglobulin tests, one had an unidentified alloantibody, and the other had an inconclusive test and developed a hemolytic transfusion reaction. Conclusion: The prevalence of clinically important red blood cell alloantibodies and hemolytic transfusion reactions among patients with unidentified alloantibodies suggests that specific laboratory techniques should be performed to identify alloantibodies in cases of pan-reactivity or autoantibodies to improve transfusion safety.