Showing 3 results for Human Immunodeficiency Virus
Mohammad Reza Amiran, Abbas Akhavan Sepahi, Rezvan Zabiollahi, Hamid Ghomi, Seyed Bahman Momen, Mohammad Reza Aghasadeghi,
Volume 4, Issue 3 (7-2016)
Abstract
Introduction: The human immunodeficiency virus (HIV) is a threat to global health and the need for finding new methods of antivirus research, in particular against HIV has increased in recent years. In this study, we investigated the ability of the cold atmospheric pressure plasma jet (CAPPJ) using helium to inhibit the replication of HIV virions. Methods: Single cycle replicable HIV (SCR-HIV) virions were produced by transfection of HEK293T cells with pmzNL4-3, pSPAX.2, and pMD2.G plasmids. The HeLa immortal cell line was infected with the virus SCR-HIV and then irradiated by CAPPJ at different voltages and times. The amount of the P24 antigen inside cell supernatant was measured by an ELISA method. Moreover, the cytotoxicity of this plasma jet and the viability of cells were evaluated by the XTT method. Results: The inhibition of HIV by CAPPJ increased with increasing of voltage and the time of radiation. The highest voltage of 12 kv at 240 s caused virus inhibition; however, the cytotoxicity of HeLa cell line also elevated with increasing of voltage and time. Conclusion: CAPPJ substantially suppresses HIV infection in vitro while causing toxicity for HeLa cell line
Elahe Akbari, Soheila Ajdari, Esmat Mirabzadeh Ardakani, Elnaz Agi, Vahid Khalaj, Azam Bolhassani,
Volume 9, Issue 2 (6-2021)
Abstract
Introduction: Recombinant subunit vaccines have been explored against various human pathogens, however, developing an effective therapeutic toward human immunodeficiency virus (HIV) infection has been challenging. So far, several recombinant HIV-1 antigens have been produced and examined for activation of desired immune responses. This study aimed to express an HIV-1 multiepitope protein as an antigen candidate to develop a vaccine. Methods: In this study, the codon-optimized encoding sequence of the designed multi-epitope construct (Gag-Pol-Env-Nef-Rev) was synthesized and subcloned into the pET-24a (+) expression vector. Then, expression of the target antigen was evaluated in E. coli BL21 (DE3) and Rosetta strains under different conditions (temperature, optical density/ OD600, isopropyl β-D-1-thiogalactopyranoside (IPTG) concentration, and time). Finally, the expression of the Gag-Pol-Env-Nef-Rev multi-epitope protein was confirmed using SDS-PAGE and western blot analysis. Results: The highly conserved and immunodominant T-cell epitopes of HIV-1 Gag, Pol, Env, Nef, and Rev proteins were used to prepare a novel Gag-Pol-Env-Nef-Rev multi-epitope construct. The gag-pol-env-nef-rev gene was successfully sub-cloned in pET-24a (+) vector and subsequently expressed in BL21 (DE3) E. coli strain under optimized conditions (1 mM IPTG, 16 h post-induction, OD 600= 0.6, and 37ºC). A clear band of ~ 35 kDa was detected by western blotting using an anti-His antibody, indicating the successful expression of our target multi-epitope protein. Conclusion: Expression of the recombinant HIV-1 multi-epitope protein was optimized in a bacterial system. The expressed protein will be purified to use as a multi-epitope protein vaccine candidate in the future.
Pratibha Shamanna, Muralidhara Sethumadhavan,
Volume 12, Issue 1 (3-2024)
Abstract
Introduction: Pneumocystis jirovecii pneumonia (PCP) remains a significant cause of pneumonia among immunocompromised individuals, despite a decline in prevalence with the advent of antiretroviral therapy (ART) for Human Immunodeficiency Virus (HIV). This study aimed to evaluate and compare the diagnostic accuracy of four distinct staining techniques for PCP in respiratory specimens. We assessed the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of these techniques against the gold standard Gomori Methenamine Silver stain (GMS), in order to identify the most effective method for diagnosing PCP. Methods: In a prospective observational study, we collected induced sputum (IS) and BAL samples from 100 immunocompromised patients and examined them microscopically for P. jirovecii cysts. We employed four staining methods for detection: Calcofluor White, Modified Toluidine Blue, Wright's stain, and Gomori Methenamine Silver stain. Results: The combination of Modified Toluidine Blue, Calcofluor White, and Wright's stains detected P. jirovecii cysts in 5% of the study population. The sensitivity of the staining methods was: 80% for Modified Toluidine Blue, 40% for Calcofluor White, and 20% for Wright's, compared to the Gomori Methenamine Silver (GMS) stain, which was used as the gold standard. All the staining methods exhibited equivalent specificity (100%). Conclusion: The Modified Toluidine Blue stain is a viable alternative to the Gomori Methenamine Silver stain due to its simplicity, speed, and applicability in resource-limited settings. The low prevalence of P. jirovecii in this study population suggests that routine cotrimoxazole prophylaxis may be effective in reducing the incidence of P. jirovecii pneumonia among HIV patients.
