High Discordance in Plasma and Genital Tract HIV-1 Drug Resistance in Indian

Women Failing First-line Therapy

MOPDA0106

S. Saravanan, PhD

Session Code: MOPDA01

Molecular Techniques of HIV-1 Analysis

21 July 2014

It is vital to investigate tissues and compartments other than blood for two important reasons (Cu-Uvin S., et al., 2010).

• From a patient perspective, it is important to determine whether antiretroviral therapy can reduce viral load in non-blood compartments.

• From a public health perspective, it is critical to know the factors that contribute to the “infectiousness” of an individual in order to devise strategies to reduce the likelihood of transmission.

Background:

Much less work has been directed at HIV in non-blood compartments and those compartments may be the potential sanctuary sites harboring HIV and impacting both the transmission and pathogenesis of HIV infection.

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Sequences were aligned (ClustalX) to an Indian subtype C reference (C.IN.AFo67155) and examined for HIV-1 subtype (REGA v2), nucleotide diversity (Highlighter HIV LANL, SLAC in HyPHY) and drug resistance associated mutations (IAS-USA and Stanford HIV Resistance Database).

Materials and Methods:

HIV-infected women (n=200) at YRG CARE in Chennai, India, who were adherent on >6 months of first-line antiretroviral therapy were enrolled.

Genital tract (2 Sno-strips in 500uL of NASBA buffer) RNA levels measured using COBAS® AMPLICOR HIV-1 MONITOR Test, v1.5 & Pol genotyping (Saravanan et al., 2009) was conducted in paired detectable samples from both compartments.

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Demographic details of enrolled study subjectsResults:

Patient Characteristics Women on First-line ART (n=200)

Viremic (n=73) Non-Viremic (n=127)

Age(Mean) years 33.8±6.3 33.4±5.2

PVL (Median) log copies/mL 4.6 (3, 5.9) Not Detected

CD4(Median) cells/µL 246 (15, 832) 530 (27, 1182)

Duration on HAART(Median) Months 35 (7, 114) 34 (6, 122)

NVP 44 (60%) 87 (68.5%)

D4T/AZT 55 (75%) 114 (90%)

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STUDY SUBJECTS ENROLLED; n = 200

VIREMIC; n=73 (36.5%) NON-VIREMIC; n=127 (63.5%)

DETECTABLE GVLn=30/42 (71%)

UNDETECTABLE GVL n=12/42 (29%)

GVL >2000 Copies/mLn=21/30 (70%)

GVL <2000 Copies/mLn=9/30 (30%)

PVL >3000 Copies/mL;n=42/73 (57.5%)

PVL <3000 Copies/mL;n=31/73 (42.5%)

Results:

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Genital and Plasma Sequences (n = 21)

Concordant mutation patterns; n = 4/21 (19%)

Discordant mutation patterns; n = 17/21 (81%)

Total patients with additional mutations in genital tract; n=11/21 (52%)

35%

24%

41%

0%

10%

20%

30%

40%

50%P

erc

en

t o

f s

ub

jec

ts

Additional inGenital

Additional inPlasma

Diverse in both

Discordance Pattern

5 NRTI – T215F, M41L, D67DN, K70T, K219E 7 NNRTI- K103E/N, H221Y, V106M, Y188H, E138A, Y181C

Results:

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Results: p = <0.005

r2 = 0.872 (p=ns)

Figure 1a: Pearson’s rank correlation for comparison between PVL and GVL in patients with discordance (n=17). 1b: Fisher’s exact test for comparison of patients with detectable and undetectable viral load in plasma and genital compartments (n=42)

95% have monophyletically clustered with Indian subtype C with one sequence clustered to CRF_02 AE

Figure 4: Phylogenetic analyses of RT

Women with detectable PVL tend to shed virus in genital secretions (p<0.005)

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Results:

Figure 6: Comparison between the presence of intermediate to high-level resistance in plasma and genital secretions in patients with discordant mutations (n=5).

Genital discordant mutations were responsible for an increase to a predicted intermediate or high level drug resistance to at least one drug in 24% of women

Figure 5: Prevalence of various NRTI and NNRTI DRMs in plasma and genital tract.

High prevalence of M184V in both compartments followed by TAMs.

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If confirmed, GVL and resistance monitoring may need to be considered to prevent sexual and perinatal HIV-1 resistance transmission in countries like India where sexual transmission is the major mode of HIV infection.

Conclusion:

High resistance discordance between plasma and the genital tract among South-Indian women failing first-line antiretroviral therapy, suggesting compartmentalization and independent viral evolution.

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Acknowledgement

IAS International scholarship (S12858)

S. Gomathi, M.ScS. Sivamalar, M.ScG. Kausalya, M.ScP. Selvamuthu, MBBSN. Kumarasamy, MBBS, PhD P. Balakrishnan, PhDSuniti Solomon, MD

Susan Cu-Uvin, MD Rami Kantor, MD

Sunil S. Solomon, MPH, PhD

Indian Council of Medical Research (ICMR) under U.S.-India Collaborative Research Supplement # Indo-US/35/2007-ECD-II

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