Imbalance of T Helper and Regulatory Cells in Patients with Tuberculosis Co-infection Acquired Immunodeficiency Syndrome

Research Article

Austin J HIV/AIDS Res. 2022; 8(1): 1050.

Imbalance of T Helper and Regulatory Cells in Patients with Tuberculosis Co-infection Acquired Immunodeficiency Syndrome

Alimu Ayiguli1#, Li Li1,2,3#*, Zhong Xuemei2#, Gong Hui1, Abudureheman Zulipikaer1, Yilamujiang Subinuer1, Liu Ya1 and Hairoula Dilare1

1Department of Clinical Research Center of Infectious Diseases (Pulmonary Tuberculosis), First People’s Hospital of Kashi, Xin Jiang, China

2Department of Respiratory and Critical Care Medicine, First People’s Hospital of Kashi, Xin Jiang, China

3State Key Laboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases in Central Asia, Xinjiang Medical University, China

#All authors contributed equally to this work

*Corresponding author: Li Li, The First People’s Hospital of Kashi. No. 120, Yingbin Avenue, Kashi, Xin Jiang, China

Received: April 08, 2022; Accepted: June 10, 2022; Published: June 17, 2022

Abstract

Background: To ensure that patients with Acquired Immunodeficiency Syndrome (HIV) and active tuberculosis(TB) co-infection (HIV-TB) can get timely treatment, we investigated T helper cells17 (Th17), Tregulatorycells(Treg) levels in the peripheral blood mononuclear cells (PBMCs) of the TB, HIV, HIVTB patients provide a better consult for prediction of HIV-TB patients.

Methods: A total of 62 patients with TB, HIV, HIV-TB and 30 healthy peoples were included in our study. The PBMCs were isolated from the peripheral blood and the level of Th17 and Treg cells were tested by flow cytometry. Serum levels of cytokines Interleukin 17 (IL-17), Interleukin 6 (IL-6) and Interleukin 10 (IL-10) were determined by ELISA.

Results: Th17/Treg imbalance in HIV-TB patients, and is lower than HIV, TB and control group (Th17/Treg: 0.10±0.03 vs 0.25±0.08 vs 0.47±0.14 or 1.03±0.47, p<0.05). The expression of IL-10 in HIV-TB patients were significantly increased than HIV, T Band control groups, IL-17 and IL-6 were significantly decased (IL-10: 28.8±4.3 vs 23.6±3.6 vs 18.5±2.9 or 14.3±1.8pg/mL, p<0.05; IL- 17: 17.1±3.6 vs 24.2±4.2 vs 28.3±5.8 or 40.9±6.8pg/mL, p<0.05; IL-6: 15.9±2.1 vs 23.8±3.4 vs 27.8±4.3 or 32.5±4.6pg/mL, p<0.05).

Conclusions: These studies revealed that Th17/Treg in HIV-TB patients was imbalanced, and suppressor T cell subsets are dominant, thus participate in immune pathogenesis of HIV-TB.

Keywords: Co-Infection AIDS with Tuberculosis; T helper Cells; Tregulatorycells; Interleukin; PBMC

Introduction

Pulmonary tuberculosis is common opportunistic infectious diseases in AIDS patients, which enable to patients, obtain dual infections of AIDS and pulmonary tuberculosis [1]. The incidence of HIV-TB patients is 30 times than normal population [2], and the risk of death is 2.87 times than ordinary tuberculosis patients [3]. CD4+ T are command center of immune system, and T helper cell (Th17) and T regulatory cell (Treg) are a subset of CD4+ T cells. Th17 mainly promotes the immune response, while Treg cells mainly suppress the immune response; they can transform each other during immune activities, there by maintaining the body’s immune balance [4-5].

Th17 cells are considered to have a significant inhibitory effect on the replication and expansion of HIV [6]. The main role of Treg is to inhibit the effect of T lymphocytes to prevent excessive autoimmune symptoms, thereby reducing the body resistance to external pathogens [7]. The Th17/Treg ratio is relatively stable under normal circumstances, but inflammation and other immune conditions can disrupt this balance [8]. For example, in the process of inflammation, TGF-β can promote the production of Treg cells and promote the differentiation of Th17 and other cells. In the late stage of inflammation, TGF-β can inhibit the immune response by inhibiting the proliferation of Treg cells, there for, the Th17/Treg balance is essential for maintaining normal immune function [9].

The importance of pro inflammatory cytokines, in support of HIV virus replication at sites of TB has been well established. Transcriptional activation of HIV virus by pro inflammatory cytokines TNF-a, interleukin (IL)-1 β, IL-6, and IL-8 is based on induction of nuclear factors [10]. IL-17 is the main effector cytokine of Th17, which can stimulate T cell activation, stimulate epithelial cells, endothelial cells and fibroblasts to produce a variety of cytokines, such as IL-6, IL-8, etc., thereby resisting extracellular bacteria, etc. Pathogen [11]. Interleukin 17 (IL-17) promotes cytokines and plays an important role in the inflammatory immune response.

There have been studies on the changes of Th17 and Treg cells in HIV/TB patients, but there are few studies on HIV-TB patients. Therefore, this study used flow cytometry and ELISA to detect the expression of Th17, Treg, CD4+ T cells and related cytokines in the peripheral blood of HIV-TB patients for detailed analysis.

Materials and Methods

Study Participants

From January 2021 to August 2021, patients with a first diagnosis of HIV-TB, TB, and HIV at the First People’s Hospital of Kashi were recruited for the study along with healthy controls. Patients and controls were matched for age, sex, and body mass index (BMI). All subjects were required to complete a questionnaire regarding basic information and provide 5mL of peripheral blood for testing. Written informed consent was obtained from all participants. Inclusion criteria: (1) it matches the diagnostic criteria of the AIDS diagnosis and treatment guidelines formulated by the Chinese Medical Association, and has undergone clinical diagnosis and laboratory tests to confirm HIV antibody positive. Some individuals were on anti-retroviral treatment (2) it matches the diagnostic criteria for tuberculosis revised and issued in 2001, and is accompanied by significant Signs of tuberculosis poisoning, the tuberculosis culture was positive, and the imaging examination was confirmed as pulmonary tuberculosis. The actuve TB patients are being treated with drugs (3) the control group were Healthy people, which HIV seronegative and mycobacterium tuberculosis culture negative. (4) Non-pregnant patients; (5) Patients older than 15 years old.

Exclusion criteria

(1) Used glucocorticoids, Immunosuppressants; (2) Other immune system diseases and other chronic diseases.

Flow Cytometric Detection of Th17 and Treg Cells

Whole blood was diluted with an equal volume of PBS before layering on Ficoll-Paque PLUS in a 15mL tube. Tubes were then centrifuged or 25 min with the brakes off at 950g PBMCs were collected from the layer between the plasma and Ficoll liquid and washed twice with PBS. Cells were pelleted by centrifugation at 350g for 7 min and then re-suspended in complete PBS. Th17 and Treg cell antibodies were respectively added into PBMC and incubate at room temperature for 30 min in the dark. After washing twice with PBS, re-suspend in 500 μL PBS and transfer to flow tube for detection by flow cytometry. According to flow cytometry results, the percentage of Th17 and Treg cells among the groups was compared. Following antibodies for Flow Cytometry: TL17-PE (Bio Legend Inc. #317306), Foxp3-PE (Bio Legend Inc. #980804). Main instruments: Flow cytometry, biological safety cabinet, low-temperature and high-speed centrifuge (Thermo Fisher Scientific, USA).

IL-6, IL-10, IL-17 Expression Level Detection

Take 2mL of fasting venous blood from the patient, centrifuge (1000×g) and take the upper serum, and detect the expression of IL- 17, IL-6 and IL-10 in serum by ELISA. The specific operation is in accordance with the instructions of the ELISA kit. IL-17, IL-6 and IL-10 Enzyme-Linked Immunosorbent Assay (ELISA) kits (Jianglai Biotechnology Company, Shanghai, China).

Statistical Analysis

SPSS 21.0 software (IBM Corp., Armonk, NY, USA) was used for statistical analyses. Quantitative data following a normal distribution are expressed as the mean ±SD. Data that had homogeneity of variance were compared by analysis of variance between multiple groups, followed by pair wise comparisons. The enumeration data was evaluated by chi-square test (Χ2).The correlation between Th17/Treg and cytokines are use person related. According to the least significant difference test, P<0.05 was considered statistically significant. Bar graphs were generated using Graph Pad Prism 8.0 (Graph Pad Software, Inc., San Diego, CA, United States).

Results

Basic Information of the Study Participants

A total of 92 people were invited to participate in this research (Table 1). There were 24 patients in TB group, 21 patients in HIV group, 17 patients in HIV-TB group, and 30 healthy persons in control group. The gender, BMI, smoking status, job occupation and age of TB, HIV and HIV-TB patients were not statistically significant compared with control group (P>0.05).