Interleukin-28B gene polymorphisms do not influence the susceptibility to HIV-infection or CD4 cell decline
Rallon, Norma Ia; Restrepo, Claraa; Naggie, Susannab; Lopez, Mariolaa; del Romero, Jorgec; Goldstein, Davidd; McHutchison, Johnb; Soriano, Vincenta; Benito, Jose Ma
aHospital Carlos III, Madrid, Spain
bDuke Clinical Research Institute, Durham, North Carolina, USA
cCentro Sanitario Sandoval, Madrid, Spain
dInstitute for Genome Sciences and Policy, Durham, North Carolina, USA.
Received 16 August, 2010
Revised 5 October, 2010
Accepted 20 October, 2010
Correspondence to Dra Norma I. Rallón, Infectious Diseases Department, Hospital Carlos III, Calle Sinesio Delgado 10, Madrid 28029, Spain. Tel: +34 91 4532500; fax: +34 91 7336614; e-mail: email@example.com
The critical role of interleukin-28B (IL28B)/interferon-λ3 (IFN-λ3) polymorphisms on the susceptibility to hepatitis C virus infection and the response to peginterferon–ribavirin therapy has encouraged exploration of similar effects on other viruses. Given that IFN-λ mediates anti-HIV-1 activity, the protective role of IL28B polymorphisms was examined in 29 seronegative individuals at risk for HIV-infection and in 68 HIV-positive carriers with and without rapid progression of immunodeficiency. No protective role of IL28B polymorphism was found examining both HIV-disease progression and HIV-protection.
Interleukin-28B (IL28B)/interferon-λ3 (IFN-λ3) belongs to the IFN type III family, shares functional characteristics with IFN type I  and induces in-vitro protection against hepatitis C virus (HCV) infection . Although several studies have demonstrated a role for single nucleotide polymorphisms (SNPs) nearby the IL28B gene on the spontaneous clearance [3,4], as well on the treatment-induced elimination [4–8] of HCV, it is currently unknown whether IL28B polymorphisms might similarly influence the outcome of other viral infections.
Although, in HIV-1, there is no evidence for spontaneous or treatment-induced cure, a subset of infected individuals, named long-term nonprogressors (LTNPs), demonstrate an intrinsic ability to control HIV-1 replication and defer immunodeficiency in the absence of antiretroviral therapy . Moreover, evidence exists on another subset of individuals who despite being exposed to HIV remain persistently HIV-negative (exposed seronegative, ESN) . A number of studies have been conducted in these special groups to ascertain whether potential immune mechanisms might account for their protection against HIV infection and/or disease progression [10,11]. Given the in-vitro antiviral effect of IFN-λ on HIV replication , it is important to examine the role of IL28B polymorphisms on HIV infection.
Separate cross-sectional case–control studies were conducted in two different well characterized cohorts of patients. The first cohort ‘HIV disease progression’ included 68 individuals with chronic HIV infection in regular follow-up at Hospital Carlos III, Madrid. The second cohort ‘HIV protection’ included 29 HIV-serodiscordant sexual couples who attended Centro Sandoval, a sexually transmitted diseases clinic located in Madrid. To participate in the study, written informed consent for genetic testing was obtained from all individuals. The study protocol was evaluated and approved by the hospital ethics committee. Both plasma and peripheral blood mononuclear cells were drawn and stored for all patients.
IL28B rs12979860 SNP genotyping was conducted on DNA specimens using the 5' nuclease assay with allele-specific TaqMan probes (ABI TaqMan allelic discrimination kit, Applied Biosystems, Carlsbad, California, USA) and the ABI7900HT Sequence Detection System (Applied Biosystems) . The main characteristics of the study population and the different parameters evaluated are expressed as median (interquartile range). Comparisons between groups were carried out using the χ2-test or Fishers's exact test, as appropriate. All statistical analyses were performed using the SPSS software version 13 (SPSS Inc., Chicago, Illinois, USA). All P-values were two-tailed, and were considered as significant only when below 0.05.
The HIV disease progression cohort included 30 LTNPs (cases) and 38 typical progressors (controls). All LTNPs were infected for more than 15 years, had plasma HIV-RNA values below 10 000 copies/ml and always had CD4 cell counts above 500 cells/μl in the absence of highly active antiretroviral therapy. Typical progressor patients were naive to antiretroviral therapy and their median CD4 cell counts were 422 (238–480) cells/μl. Overall, 57% were co-infected with HCV (73% of LTNPs vs. 45% of typical progressors).
The HIV protection cohort  included 29 HIV-ESN individuals (cases) and 29 HIV-infected partners (controls). Median CD4 cell count in HIV-infected partners was 496 (207–705) cells/μl and length of infection was 108 (30–188) months. At inclusion, 65% (19/29) of them had undetectable plasma viremia, ranging in the remaining 10 patients from 68 to 238 131 HIV-RNA copies/ml. Thirty-eight percent of ESN were co-infected with HCV. Fifteen (52%) ESN admitted never using condoms, nine (31%) sporadic use, and five (17%) frequent use. Considering risk behaviours and viral load of HIV-infected partner, ESN were divided into those with low (19; 65%) or moderate-high (10; 35%) level of HIV exposure.
Both cohorts, HIV disease progression and HIV protection, were in Hardy–Weinberg equilibrium, with a C allele frequency of 70 and 72%, respectively. The prevalence of rs12979860 genotypes in the HIV disease progression cohort was CC 50%, CT 40%, and TT 10%. The distribution of IL28B polymorphisms was similar in LTNPs and typical progressors (CC genotype was present in 54 and 47%, respectively, P = 0.63; Table 1). The prevalence of IL28B rs12979860 genotypes in the HIV protection cohort was CC 54%, CT 36%, and TT 10%. Although the distribution of the different IL28B genotypes did not differ significantly between ESN and their HIV-positive partners, the prevalence of the CC genotype tended to be greater in ESN than in their HIV-positive partners (62 vs. 45%, respectively; P = 0.19; Table 1).
The IL28B rs12979860 SNP is one of the most important discoveries of a genetic trait associated with control of a chronic viral infection. The fact that this SNP is located 3 kb upstream the IL28B/IFN-λ3 gene  and that IFN-λ is known to mediate anti-HIV-1 activity  might provide the basis for implicating a role of innate immunity against both HCV and HIV infections. Our results, however, do not support a role for this IL28B SNP in the protection from HIV disease progression in infected persons, as the prevalence of the protective genotype (CC) was similar in LTNPs and typical progressors. Likewise, the IL28B SNP does not seem to confer any protection against HIV acquisition, as the prevalence of the CC genotype did not differ significantly in ESN and their HIV-positive partners.
IFN-λ exerts bioactivities that overlap those of type I IFNs, such as antiviral activity  and upregulates the intracellular expression of type I IFNs and APOBEC3G/3F, a well known anti-HIV-1 cellular factor . Thus, it is somewhat unexpected that the IL28B SNP did not significantly influence HIV control, whereas the effect has shown to be quite strong on HCV infection. Therefore, large differences in the control of HCV and HIV infections may exist. The biological mechanisms involved in the association of the IL28B SNP over HCV control are still unclear. Although the CC genotype might modulate the regulation of IL28B protein expression, and in this way influence innate immunity, in spite of the previously reported IFN-λ-mediated anti-HIV-1 activity , we did not find any significant association between IL28B polymorphisms and HIV disease progression nor HIV protection. Thus, the biological mechanisms underlying the association of the IL28B SNP with HCV control could be other than the direct IL28B-mediated antiviral activity. Alternatively, different mechanisms might be operating in vivo in the control of HCV and HIV infections.
This work was supported in part by grants from Fundación Investigacion y Educacion en SIDA (IES), Red de Investigacion en SIDA (RIS, FIS-RD06/0006), Agencia Lain Entralgo and the European Union 6th Framework Programme (NEAT, LSHP-CT-2006- 037570).
The authors would like to thank all patients who participated in the study.
N.I.R., V.S., J.M.B., S.N., and J.McH. designed the study. N.I.R., C.R., J.M.B., and M.L. did the virological studies and collected the specimens. N.I.R., C.R., D.G. and J.McH. performed SNP genotyping. V.S. and J.R. were responsible for and analysed the demographics, clinical and therapeutic information of the study population. N.I.R., J.M.B., V.S., S.N. and J.McH. wrote the manuscript draft. All authors revised and approved the final submission.
1. Kotenko SV, Gallagher G, Baurin VV, Lewis-Antes A, Shen M, Shah NK, et al. IFN-lambdas mediate antiviral protection through a distinct class II cytokine receptor complex. Nat Immunol 2003; 4:69–77.
2. Marcello T, Grakoui A, Barba-Spaeth G, Machlin ES, Kotenko SV, MacDonald MR, et al. Interferons alpha and lambda inhibit hepatitis C virus replication with distinct signal transduction and gene regulation kinetics. Gastroenterology 2006; 131:1887–1898.
3. Thomas DL, Thio CL, Martin MP, Qi Y, Ge D, O'Huigin C, et al. Genetic variation in IL28B and spontaneous clearance of hepatitis C virus. Nature 2009; 461:798–801.
4. Rallón NI, Naggie S, Benito JM, Medrano J, Restrepo C, Goldstein D, et al. Association of a single nucleotide polymorphism near the interleukin-28B gene with response to hepatitis C therapy in HIV/hepatitis C virus-coinfected patients. AIDS 2010; 24:F23–F29.
5. Ge D, Fellay J, Thompson AJ, Simon JS, Shianna KV, Urban TJ, et al. Genetic variation in IL28B predicts hepatitis C treatment-induced viral clearance. Nature 2009; 461:399–401.
6. Suppiah V, Moldovan M, Ahlenstiel G, Berg T, Weltman M, Abate ML, et al. IL28B is associated with response to chronic hepatitis C interferon-alpha and ribavirin therapy. Nat Genet 2009; 41:1100–1104.
7. Tanaka Y, Nishida N, Sugiyama M, Kurosaki M, Matsuura K, Sakamoto N, et al. Genome-wide association of IL28B with response to pegylated interferon-alpha and ribavirin therapy for chronic hepatitis C. Nat Genet 2009; 41:1105–1109.
8. Rauch A, Kutalik Z, Descombes P, Cai T, Di Iulio J, Mueller T, et al. Genetic variation in IL28B is associated with chronic hepatitis C and treatment failure: a genome wide association study. Gastroenterology 2010; 138:1338–1345.
9. Pantaleo G, Menzo S, Vaccarezza M, Graziosi C, Cohen OJ, Demarest JF, et al. Studies in subjects with long-term nonprogressive HIV infection. N Engl J Med 1995; 332:209–216.
10. Kulkarni P, Butera S, Duerr A. Resistance to HIV-1 infection: lessons learned from studies of highly exposed persistently seronegative (HEPS) individuals. AIDS Rev 2003; 5:87–103.
11. Piacentini L, Biasin M, Fenizia C, Clerici M. Genetic correlates of protection against HIV infection: the ally within. J Intern Med 2009; 265:110–124.
12. Hou W, Wang X, Ye L, Zhou L, Yang ZQ, Riedel E, et al. Lambda interferon inhibits human immunodeficiency virus type 1 infection of macrophages. J Virol 2009; 83:3834–3842.
13. Livak K. Allelic discrimination using fluorogenic probes and the 5' nuclease assay. Genet Anal 1999; 14:143–149.
14. Restrepo C, Rallón NI, del Romero J, Rodríguez C, Hernando V, López M, et al. Low-level exposure to HIV induces virus-specific T cell responses and immune activation in exposed HIV-seronegative individuals. J Immunol 2010; 185:982–989.
This article has been cited 4 time(s).
Journal of Viral HepatitisValue of IL28B genotyping in patients with HCV-related mixed cryoglobulinemia: results of a large, prospective studyJournal of Viral Hepatitis
AIDS Research and Human RetrovirusesIL28B CC Genotype Is Associated with Higher All-Cause Mortality in Antiretroviral-Treated HIV-Infected PatientsAIDS Research and Human Retroviruses
Bmc Infectious DiseasesLack of evidence to support the association of a single IL28B genotype SNP rs12979860 with the HTLV-1 clinical outcomes and proviral loadBmc Infectious Diseases
Journal of Infectious DiseasesIL28B Single-Nucleotide Polymorphism rs12979860 Is Associated With Spontaneous HIV Control in White SubjectsJournal of Infectious Diseases
© 2011 Lippincott Williams & Wilkins, Inc.