HIV-1 Subtype F in Single and Dual Infections in Puerto Rico: A Potential Sentinel Site for Monitoring Novel Genetic HIV Variants in North America.

To the Editor: Although international efforts to systematically collect, characterize, and classify HIV isolates from around the world have increased considerably, data on HIV-1 genetic variations in Puerto Rico are limited. This island (population 3.7 million) has one of the highest AIDS incidence rates in the United States (53.3 cases per 100,000) (1). To evaluate the potential for a multiple subtype distribution pattern in Puerto Rico, we analyzed genetic variations between HIV-1 strains isolated from peripheral blood mononuclear cells of 63 asymptomatic HIV-infected female commercial sex workers from 12 communities. These participants were part of 290 female commercial sex workers followed in a larger cross-sectional study of risk behavior (2).

HIV-1 subtypes F (n = 4) and B (n = 44) strains were identified in persons infected with a single viral subtype with a molecular screening assay based on restriction fragment length polymorphism (RFLP) analysis and with DNA sequencing of the viral protease gene-prot (3). The remaining 15 specimens were classified by RFLP as potential dual infections. Further cloning and sequencing of prot from three of these specimens confirmed one dual infection involving subtypes F and B viruses and identified two infections caused by genetically distinct quasispecies of subtype B variants.

In further detailed pairwise analysis of HIV-1 prot genes, a small nucleotide divergence of 0.3% (0.0 to 1.1) within subtype F contrasted with a typical value of 6.3% (5.1 to 7.8) for the intrasubtype distance within subtype B prot sequences (4). The 99% similarity between prot subtype F Puerto Rican sequences suggested an epidemiologic link or a recent introduction of subtype F in Puerto Rico. Comparative sequence analysis of the C2-V3 env is useful in establishing the time that elapsed from infection on the basis of an annual nucleotide divergence of 0.5% to 1% in this region (5). Such analysis has been used to study the epidemiologic link between cases (4,6). Thus, we compared env sequences from two of five persons infected with prot subtype F strains. This analysis provided several observations. Env nucleotide divergence of 13.2% did not support a direct epidemiologic link between these strains. Furthermore, the relatively high intrasubtype diversity between env sequences suggested that evolution from a common progenitor would have taken a minimum of approximately 13 years. Phylogenetic analysis classified these two env sequences as subtype B, indicating that at least some of Puerto Rican prot subtype F viruses represent HIV-1 mosaics involving closely related prot F and significantly divergent env B sequences. Overall, discrepancy in both subtype assignment and nucleotide diversities within prot and env regions may indicate that distinct F/B mosaics circulating in Puerto Rico were likely the result of recombination between highly homogeneous subtype F of relatively recent arrival and divergent resident subtype B viruses.

HIV-1 infections with subtype F strains including B/F mosaics have been reported in Brazil (3,7). To evaluate a potential HIV-1 linkage between Brazil and Puerto Rico, a comparative phylogenetic analysis was done on subtype F viral prot sequences from these countries. This analysis documented that HIV-1 subtype F strains in Puerto Rico are distinct from both Brazilian and Romanian viruses. Furthermore, our results show that genetic analysis of prot allows tracking of subtype F viruses of different origin. Recently, by this approach, HIV-1 prot subtype F of Puerto Rican origin and F prot/B env mosaic were identified in HIV-1-infected persons in New York city (8). Observation of HIV-1 subtype F strains in Puerto Rico together with the recent report describing the first cases of such infections in New York indicates the potential for further emergence of subtype F on the North American continent. The presence of a complex distribution pattern of subtype F infections in Puerto Rico has serious implications for the evaluation and development of HIV diagnostics and vaccines.

Supported in part by grant G12RR-03050 (Y.Y.).

The nucleotide HIV-1 sequences obtained in this study were submitted to GenBank; their accession numbers are AF096813-AF096833.

References

(1.) Centers for Disease Control and Prevention. HIV/AIDS surveillance report no. 2; 1997;9.

(2.) Drugs, HIV infection and risk behaviors among Puerto Rican sex workers, 1994-1996. Grant: NIAID/RCMI #G12RR03051 [Dr. Margarita Alegria]. Sociomedical Research Department, Graduate School of Public Health, University of Puerto Rico, Medical Sciences Campus, San Juan, Puerto Rico.

(3.) Ramos A, Tanuri A, Schechter M, Rayfield MA, Hu DJ, Cabral MC, et al. HIV-1 dual infections are an integral part of the HIV epidemic in Brazil. Emerg Infect Dis 1999;5:65-74.

(4.) Janini LM, Tanuri A, Schechter M, Peralta JM, Vicente AC, De la Torre N, et al. Horizontal and vertical transmission of human immunodeficiency virus type 1 dual infections caused by viruses of subtypes B and C. J Infect Dis 1998;177:227-31.

(5.) Myers G, Korber B, Berzofski JA, Smith RF, and Database and Analysis Staff, editors. Human retroviruses and AIDS 1991: a compilation and analysis of nucleic acid and amino acid sequences. Los Alamos (NM): Los Alamos National Laboratory; 1991.

(6.) Ou CY, Ciesielski C, Myers G, Bandea CI, Luo CC, Korber BT, et al. Molecular epidemiology of HIV transmission in a dental practice. Science 1992;256:1167-71.

(7.) Sabino EC, Shpaer EG, Morgado MG, Korber BT, Diaz RS, Bongertz V, et al. Identification of human immunodeficiency virus type 1 envelope genes recombinant between subtypes B and F in two epidemiologically linked individuals from Brazil. J Virol 1994;68:6340-6.

(8.) Weidle PJ, Ganea CE, Pieniazek D, Ramos CA, Ernst JA, McGowan JP, et al. Prevalence of HIV-1 group M, non-B-subtypes in Bronx, New York community: a sentinel site for monitoring of HIV genetic diversity in the United States. In: Proceedings of the 12th World AIDS Conference; 1998 Jun; Geneva, Switzerland [abstract no. 13225].

Idhaliz Flores,(*) Danuta Pieniazek,([dagger]) Nitza Moran,(*) Angel Soler,(*) Nayra Rodriguez,(*) Margarita Alegria,([double dagger]) Mildred Vera,([double dagger]) Luiz M. Janini,([dagger]) Claudiu I. Bandea,([dagger]) Artur Ramos,([dagger]) Mark Rayfield,([dagger]) and Yasuhiro Yamamura(*)

(*) Ponce School of Medicine, Ponce, Puerto Rico; ([dagger]) Centers for Disease Control and Prevention, Atlanta, GA; ([dagger]) University of Puerto Rico, San Juan, Puerto Rico