Distinct human immunodeficiency virus strains in the bone marrow are associated with the development of thrombocytopenia

Abstract

We analyzed bone marrow and blood from human immunodeficiency virus type 1 (HIV-1)-infected individuals and described the HIV-1 quasispecies in these cellular compartments. HIV-1 isolates from the bone marrow of thrombocytopenic patients contained distinct amino acids in the V3 loop and infected T-cell lines, implicating this virus in the development of thrombocytopenia.

FIG. 1

Alignment of V3 loop amino acid sequences of HIV-1 from bone marrow (A) and blood (B). The most prominent sequence for each patient is shown, using the one-letter amino acid code. The ratio to the right indicates the frequency of the sequence in each patient’s bone marrow or blood sample. Identical amino acids and deletions are represented by dots and dashes, respectively. Amino acid insertions are shown where they occur, but dashes are introduced into the rest of the sequences to optimize alignment. Boldfaced amino acids designate positions 11 and 13 of the V3 loop. Asterisks indicate the V3 loop sequences used to construct HIV-1 molecular clones, and the numbers next to them indicate the corresponding molecular clones. Roman numerals I, II, and III designate amino acid sequences that are identical in the bone marrow and blood of two of the patients.

FIG. 2

Rooted phylogenetic tree showing the V3 loop sequences of HIV-1 from the bone marrow of 12 patients and the blood of 3 of the patients. The scale bar indicates 5% divergence, at the nucleotide level, between any two sequences. BM, bone marrow; Bld, blood.

FIG. 3

The V3 loop consensus sequences from bone marrow HIV-1 sequences of patients with and without TP. Subscripts designate the frequency of each amino acid for every position in the V3 loop. The TP consensus was compiled from the 30 amino acid sequences of patients A to C. The non-TP consensus was compiled from the 83 amino acid sequences of patients D to L.

FIG. 4

Replication kinetics of HIV-1 isolates in activated PBMC (A), primary macrophages (B), and MT-2 cells (C). p120 and p125 are T-cell-tropic and macrophage-tropic HIV-1 molecular clones, respectively. Activated PBMC and primary macrophages correspond to donor 2 of Table 2. Supernatants were assayed for reverse transcriptase (RT) activity (expressed as counts per minute per milliliter of supernatant) every 4 days. Each point represents the average of three independent measurements.