Lentivirally overexpressed T-bet regulates T-helper cell lineage commitment in chronic hepatitis B patients

Liu,Xueni; Tang,Zhenghao; Zhang,Yi; Hu,Jianjun; Li,Dan; Zang,Guoqing; Yu,Yongsheng

doi:10.3892/mmr.2012.905

Lentivirally overexpressed T-bet regulates T-helper cell lineage commitment in chronic hepatitis B patients

Authors:
- Xueni Liu
- Zhenghao Tang
- Yi Zhang
- Jianjun Hu
- Dan Li
- Guoqing Zang
- Yongsheng Yu
View Affiliations

Affiliations: Department of Infectious Diseases, Sixth People's Hospital, Shanghai Jiaotong University, Shanghai 200233, P.R. China
Published online on: May 8, 2012 https://doi.org/10.3892/mmr.2012.905
Pages: 361-366

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Chronic hepatitis B virus (HBV) infection is commonly considered to occur as a result of disturbance of the immune system. T-box expressed in T cells (T-bet) is an essential transcription factor for T helper (Th) cell differentiation and function. The aim of this study was to investigate the effect of T-bet overexpression on Th cell differentiation and the possible mechanism in chronic hepatitis B (CHB) patients. CD4+ T cells from the peripheral blood of 23 CHB patients, 8 acute hepatitis B (AHB) patients and 10 healthy controls were isolated. T-bet mRNA expression of CD4+ T cells was detected by quantitative real-time polymerase chain reaction (PCR). The T-bet DNA fragment was subcloned into the pGC-FU vector containing GFP to generate a recombinant lentiviral vector, pGC-FU-T-bet, while a no-load pGC-FU vector was used as the negative control. After transduction into CD4+ T cells from another 22 CHB patients, the induction of Th1- and Th2-type cytokines was assayed by an enzyme-linked immunosorbent assay (ELISA), and RT-PCR and western blot analysis were used to measure the mRNA and transcription levels of H2.0-like homeobox (HLX1), GATA-3 and STAT-6. T-bet mRNA expression in CD4+ T cells from AHB patients was enhanced compared with CHB patients and healthy controls. Th1-type cytokines and HLX1 expression was upregulated, while Th2-type cytokines and GATA-3 and STAT-6 expression was repressed after lentiviral introduction of T-bet. In conclusion, lentivirally overexpressed T-bet regulates Th cell lineage commitment in CHB patients, which may be mediated by regulating HLX1, GATA-3 and STAT-6 expression.

View Figures

View References

1	Guidotti LG and Chisari FV: Immunobiology and pathogenesis of viral hepatitis. Annu Rev Pathol. 1:23–61. 2006. View Article : Google Scholar : PubMed/NCBI
2	Asabe S, Wieland SF, Chattopadhyay PK, et al: The size of the viral inoculum contributes to the outcome of hepatitis B virus infection. J Virol. 83:9652–9662. 2009. View Article : Google Scholar : PubMed/NCBI
3	Lewis MD, Miller SA, Miazgowicz MM, Beima KM and Weinmann AS: T-bet’s ability to regulate individual target genes requires the conserved T-box domain to recruit histone methyltransferase activity and a separate family member-specific transactivation domain. Mol Cell Biol. 27:8510–8521. 2007.
4	Beima KM, Miazgowicz MM, Lewis MD, Yan PS, Huang TH and Weinmann AS: T-bet binding to newly identified target gene promoters is cell type-independent but results in variable context-dependent functional effects. J Biol Chem. 281:11992–12000. 2006. View Article : Google Scholar : PubMed/NCBI
5	Pai SY, Truitt ML and Ho IC: GATA-3 deficiency abrogates the development and maintenance of T helper type 2 cells. Proc Natl Acad Sci USA. 101:1993–1998. 2004. View Article : Google Scholar : PubMed/NCBI
6	Ansel KM, Djuretic I, Tanasa B and Rao A: Regulation of Th2 differentiation and Il4 locus accessibility. Annu Rev Immunol. 24:607–656. 2006. View Article : Google Scholar : PubMed/NCBI
7	Gutiérrez-García ML, Fernandez-Rodriguez CM, Lledo-Navarro JL and Buhigas-Garcia I: Prevalence of occult hepatitis B virus infection. World J Gastroenterol. 17:1538–1542. 2011.
8	Raimondo G, Pollicino T, Cacciola I and Squadrito G: Occult hepatitis B virus infection. J Hepatol. 46:160–170. 2007. View Article : Google Scholar : PubMed/NCBI
9	Mizukoshi E, Sidney J, Livingston B, et al: Cellular immune responses to the hepatitis B virus polymerase. J Immunol. 173:5863–5871. 2004. View Article : Google Scholar : PubMed/NCBI
10	Brooks DG, Teyton L, Oldstone MB and McGavern DB: Intrinsic functional dysregulation of CD4 T cells occurs rapidly following persistent viral infection. J Virol. 79:10514–10527. 2005. View Article : Google Scholar : PubMed/NCBI
11	Jiang R, Feng X, Guo Y, et al: T helper cells in patients with chronic hepatitis B virus infection. Chin Med J (Engl). 115:422–424. 2002.PubMed/NCBI
12	Akpolat N, Yahsi S, Godekmerdan A, Demirbag K and Yalniz M: Relationship between serum cytokine levels and histopathological changes of liver in patients with hepatitis B. World J Gastroenterol. 11:3260–3263. 2005. View Article : Google Scholar : PubMed/NCBI
13	Kao C, Oestreich KJ, Paley MA, et al: Transcription factor T-bet represses expression of the inhibitory receptor PD-1 and sustains virus-specific CD8⁺ T cell responses during chronic infection. Nat Immunol. 12:663–671. 2011. View Article : Google Scholar : PubMed/NCBI
14	Kootstra NA and Verma IM: Gene therapy with viral vectors. World J Gastroenterol. 43:413–439. 2003.PubMed/NCBI
15	Montini E, Cesana D, Schmidt M, et al: The genotoxic potential of retroviral vectors is strongly modulated by vector design and integration site selection in a mouse model of HSC gene therapy. Clin Invest. 119:964–975. 2009. View Article : Google Scholar : PubMed/NCBI
16	Abordo-Adesida E, Follenzi A, Barcia C, et al: Stability of lentiviral vector-mediated transgene expression in the brain in the presence of systemic antivector immune responses. Hum Gene Ther. 16:741–751. 2005. View Article : Google Scholar : PubMed/NCBI
17	Mátrai J, Chuah MK and VandenDriessche T: Recent advances in lentiviral vector development and applications. Mol Ther. 18:477–490. 2010.
18	Meuer SC, Hussey RE, Cantrell DA, et al: Triggering of the T3-Ti antigen-receptor complex results in clonal T-cell proliferation through an interleukin 2-dependent autocrine pathway. Proc Natl Acad Sci USA. 81:1509–1513. 1984. View Article : Google Scholar : PubMed/NCBI
19	Liao W, Lin JX, Wang L, Li P and Leonard WJ: Modulation of cytokine receptors by IL-2 broadly regulates differentiation into helper T cell lineages. Nat Immunol. 12:551–559. 2011. View Article : Google Scholar : PubMed/NCBI
20	Zheng WP, Zhao Q, Zhao X, et al: Up-regulation of Hlx in immature Th cells induces IFN-gamma expression. J Immunol. 172:114–122. 2004. View Article : Google Scholar : PubMed/NCBI
21	Mullen AC, Hutchins AS, High FA, et al: Hlx is induced by and genetically interacts with T-bet to promote heritable T(H)1 gene induction. Nat Immunol. 3:652–658. 2002.PubMed/NCBI
22	Hamalainen-Laanaya HK, Kobie JJ, Chang C and Zeng WP: Temporal and spatial changes of histone 3 K4 dimethylation at the IFN-gamma gene during Th1 and Th2 cell differentiation. J Immunol. 179:6410–6415. 2007. View Article : Google Scholar : PubMed/NCBI
23	Farrar JD, Ouyang W, Löhning M, et al: An instructive component in T helper cell type 2 (Th2) development mediated by GATA-3. J Exp Med. 193:643–650. 2001. View Article : Google Scholar : PubMed/NCBI
24	Nawijn MC, Dingjan GM, Ferreira R, et al: Enforced expression of GATA-3 in transgenic mice inhibits Th1 differentiation and induces the formation of a T1/ST2-expressing Th2-committed T cell compartment in vivo. J Immunol. 167:724–732. 2001. View Article : Google Scholar : PubMed/NCBI
25	Skapenko A, Leipe J, Niesner U, et al: GATA-3 in human T cell helper type 2 development. J Exp Med. 199:423–428. 2004. View Article : Google Scholar : PubMed/NCBI
26	Zhu J, Min B, Hu-Li J, et al: Conditional deletion of Gata3 shows its essential function in T(H)1-T(H)2 responses. Nat Immunol. 5:1157–1165. 2004. View Article : Google Scholar : PubMed/NCBI
27	Hwang ES, Hong JH and Glimcher LH: IL-2 production in developing Th1 cells is regulated by heterodimerization of RelA and T-bet and requires T-bet serine residue 508. J Exp Med. 202:1289–1300. 2005. View Article : Google Scholar : PubMed/NCBI
28	Zhu J, Guo L, Watson CJ, Hu-Li J and Paul WE: Stat6 is necessary and sufficient for IL-4’s role in Th2 differentiation and cell expansion. J Immunol. 166:7276–7281. 2001.
29	Goenka S and Kaplan MH: Transcriptional regulation by STAT6. Immunol Res. 50:87–96. 2011. View Article : Google Scholar