Increased mRNA expression of key cytokines among suspected cases of Pneumocystis jirovecii infection.

BACKGROUND: Pneumocystis pneumonia (PCP) is a fatal infectious disease caused by Pneumocystis jirovecii (PJP). The major factor relevant to morbidity and mortality seems to be the host inflammatory reaction. The objective of this study was to evaluate the role of IL-2, IL-4, IL-10, and IL-13 cytokine mRNA expression among suspected P. jirovecii infection. METHODS: This was a cross-sectional analytical study undertaken in Aseer region, Saudi Arabia. One hundred suspected PCP cases and 100 healthy controls were included in the study. Basic clinical manifestations, radiological findings, microbiological and immunological findings were extracted from the hospital records from January 2019 to August 2019, Pneumocystis detection was done by immune-fluorescent staining (IFAT, Gomorimethanamine silver staining (GMSS), Giemsa staining, Toluidine blue O (TBO), and Pneumocystis RT-PCR. RESULTS: Increased more than 5 fold, 3 fold, 4 fold, and 7 fold of IL-2, IL-4, IL-10, and IL-13 mRNA expression were observed in PCP cases compared to controls. Higher expression of IL-2 mRNA was connected with crept, wheezing and chest X-ray findings like central perihilar infiltrate, patchy infiltrate, consolidation, hilar lymphadenopathy, pneumothorax, pleural effusion which showed higher expression compared to counterpart (p< 0.0001). Higher expression of IL-4 mRNA was found to be significantly associated with weight loss (p=0.002), dyspnea (p=0.003), crept (p=0.01), and chest X-ray findings (p< 0.0001). Significantly increased expression of IL-10 mRNA was observed to be associated with weight loss, dyspnea, night sweats, wheezing, and different findings of chest X-ray compared to their counterparts, whereas, IL-13 mRNA was observed in cases with fever. Suspected cases of PCP confirmed positive by IFTA with higher IL-2, IL-4, and IL-10 mRNA expression compared to negative cases. RT-PCR confirmed PCP cases had significantly higher expression of IL-2, IL-4, and IL-10 as well as IL-13 mRNA compared to negative cases. Positive detected cases by GMSS showed higher IL-2, IL-10 mRNA expression, while Giemsa showed only higher IL-4 mRNA expression compared to negative cases. CONCLUSION: Confirmed cases of P. jirovecii showed higher IL-2, IL-4, IL-10, and IL-13 mRNA expression comparatively to negative cases. Increased expression of cytokines may be indicative of infection severity and could help in patients' management.

Evaluation of modified agglutination test and enzyme-linked immunosorbent assay in detection of toxoplasma gondii antibody in human sera / Avaliação do método de aglutinação direta e do ensaio imunoenzimático na detecção de anticorpos para toxoplasma gondii em soros humanos

Toxoplasmosis is a zoonosis of high prevalence around the world, leading to serious congenital alterations and in immune compromised individuals. Its diagnosis is based in the detection of serum antibodies. This work aimed to compare serum antibody to Toxoplasma gondii detection by ELISA for IgG antibodies, and the MAT with the results of IFAT. From 73 samples examined by IgG-IFAT, 53 (72.6%) were positive, IgG-ELISA 55 (75.4%) and MAT, 58 (79.4%). No significative differences among results of tests are found. MAT shows high sensibility (100%) and lower specificity (75%), there were not false-negative results, but showed false-positives (25%). ELISA showed sensibility of 98.1% and specificity of 85,0%, false-negative rate of 1.9% and false-positive rate of 15.0%. MAT performance, when compared with IFAT, suggests the utilization of this method in diagnostic and epidemiological preliminary tests, besides ELISA has been used as a confirmatory test, for its specificity.

A toxoplasmose é uma zoonose de elevada prevalência em todo o mundo, podendo causar sérias alterações congênitas e em pacientes imunodeprimidos. Seu diagnóstico, na maioria das vezes, baseia-se na detecção de anticorpos séricos. Neste trabalho propôs-se comparar a detecção de anticorpos séricos para Toxoplasma pelo ELISA para IgG, e o MAD com os resultados da RIFI, em soros de indivíduos provenientes de um assentamento localizado em Eldorado, MS. Das 73 amostras examinadas pela RIFI-IgG, 53 (72,6%) amostras foram positivas, para o ELISA-IgG 55 (75,4%) e o MAD, 58 (79,4%). Não houve diferença significativa entre os resultados dos testes. O MAD apresentou alta sensibilidade (100%) e menor especificidade (75%), 25% de falso-positivos e sem resultados falso-negativos. Já o ELISA apresentou sensibilidade de 98,1% e especificidade de 85,0%, com taxa de falsos-negativos de 1,9% e de falso-positivos de 15,0%. A performance do MAD, quando comparado à RIFI, sugere a utilização deste método em provas de triagem diagnóstica e epidemiológica, enquanto que o ELISA seria utilizado como prova confirmatória, devido a sua especificidade.