Hepatocyte-tumor cell interaction in vitro. I. Conditions for rosette formation and inhibition by anti-H-2 antibody.

Murine hepatocytes, isolated by an in situ collagenase-perfusion technique and cultured in Petri dishes, were shown to form rosettes with liver-metastasizing syngeneic tumor cells. Pretreatment of the tumor cells with neuraminidase generally increased the binding, whereas pretreatment of the liver cells with neuraminidase abolished the binding completely. The tumor-cell binding may be mediated by the previously described lectin-like receptor of hepatocytes that also was sensitive to neuraminidase treatment and that bound desialylated cells better than normal cells. Anti-H-2 sera could efficiently inhibit the rosette formation of metastatic tumor cells with the hepatocytes, which points to a possible role of H-2 molecules in this interaction of neoplastic and normal cells.

Productive infection and cell-free transmission of human T-cell leukemia virus in a nonlymphoid cell line.

Human T-cell leukemia virus (HTLV), American PL isolate, was transmitted by cocultivation and by cell-free filtrates to a nonlymphoid human osteogenic sarcoma (HOS) cell line, designated HOS/PL, but not to nine other lines bearing receptors for HTLV. HOS and HOS/PL cells are not dependent on interleukin-2 and do not express interleukin-2 receptors that are recognized by anti-Tac monoclonal antibody. HTLV released by the Japanese MT2 cell line was also transmitted to HOS cells. The infected HOS cells release substantial titers of progeny HTLV which is antigenically indistinguishable from parental virus and is able to transform T cells.

Different patterns of lectin binding and cell surface sialylation detected on related high- and low-metastatic tumor lines.

We have analyzed cell surface-bound carbohydrates in two different model systems for metastasis composed of closely related tumor cell lines with differing metastatic potential. The first system studied was that of the DBA/2-derived T-lymphoma lines (Eb/ESb) and some recently established sublines of ESb with altered metastatic behavior (ESb-M and ESb-MR). The second system consisted of the highly metastatic MDAY-D2 cells, a wheat germ agglutinin-resistant low metastatic subline MDW40, and two metastatic revertants from the latter. The cells were stained with fluorescein isothiocyanate-conjugated lectins and analyzed by flow cytofluorography. All low-metastatic tumor lines expressed receptor sites for the lectins soybean agglutinin (SBA) and Vicia villosa (VV). The metastatic lines had the respective lectin binding sites blocked by sialic acid (SA). A good correlation was found within the cell lineages Eb leads to ESb leads to ESb-M leads to ESb-MR and MDAY-D2 leads to MDW40 leads to MDW40M1 between reactivity of SBA and VV and metastatic potential. The amount of neuraminidase-accessible SA was similar for all cell types (except MDW40) indicating differences in the positioning of SA. For high-metastatic ESb cells, the sialylation of SBA and VV receptor sites was paralleled by a relative decrease of SA associated with receptor sites for peanut agglutinin. Low-metastatic Eb cells, in contrast, had their peanut agglutinin receptor sites sialylated but expressed asialylated SBA and VV receptor sites. Eb cells were also found to have 2-fold higher activities in galactose-specific sialyltransferases. It is proposed that the differences in positioning of SA on the cell surface leading to masking or unmasking of terminal sugars could influence the metastatic potential of tumor cells.

Development of HIV-1 group-specific neutralizing antibodies after seroconversion.

OBJECTIVE: To study the induction of group-specific (gs) neutralizing antibodies to HIV-1 after seroconversion. DESIGN AND METHODS: Serum samples taken sequentially from seven Dutch homosexual men and four British haemophiliacs (anonymous sample, therefore sex not known) before and after seroconversion were tested for neutralizing antibodies effective against five diverse HIV-1 strains. Strains of HIV-1 tested included isolates from the United States, Europe and Africa. RESULTS: The gs neutralizing antibody response varied between individuals. Only five of the 11 individuals studied produced detectable neutralizing antibodies to laboratory-adapted HIV-1 strains (for example, IIIB) within 32 weeks of seroconversion. Most individuals initially produced antibodies effective against US/European isolates; the response then generally broadened to include the more diverse strains, i.e., African. CONCLUSIONS: These results suggest that the gs neutralizing target for HIV-1 is poorly immunogenic in vivo and is probably not highly conserved among diverse HIV-1 strains.

Antibodies to a putative HIV gp41 immunosuppressive peptide, pHIVIS (583-599), do not correlate significantly with outcome in HIV infection.

We have studied the prevalence of antibodies to peptides derived from the transmembrane protein of HIV, gp41. Previous work has suggested that the presence of antibodies to the gp41 peptide known as pHIVIS (env 583-599) is associated with protection from immunosuppression in HIV infection. We studied 171 sequential sera from 55 HIV-1-infected people in various clinical stages of disease. There was no significant association between antibodies to pHIVIS and clinical status in this study. Although pHIVIS has sequence similarity to the putative immunosuppressive region of the C-type oncornaviruses (p15E), antibodies to this peptide do not appear to be associated with protection from immunosuppression in natural HIV infection.

Molecular epidemiology of HIV-1 in the former Soviet Union: analysis of env V3 sequences and their correlation with epidemiologic data.

OBJECTIVE: To investigate the HIV-1 V3 sequence diversity in the former Soviet Union in 30 subjects infected with HIV-1 via different modes of transmission. PATIENTS: A cohort of children infected after exposure to nonsterile needles during the epidemic in 1988-1989 in southern Russia (Elista, n = 12 and Rostov-on-Don, n = 10), and eight HIV-seropositive subjects from Belarus (Minsk), infected via sexual (n = 7) and parenteral (n = 1) infection. METHODS: The HIV-1 V3 encoding region was amplified by nested polymerase chain reaction on DNA of primary peripheral blood mononuclear cells collected from the study subjects and then cloned and sequenced. RESULTS: The alignment of 127 V3 sequences from 22 patients in the cohort group demonstrated common consensus sequences in both the Elista and Rostov samples. The average means of interperson variation were 5.9 and 6.6% in Elista and Rostov subjects, respectively, and comparable to the mean intraperson variation. The average mean interperson variation between nucleotide sequences of HIV patients infected through sexual transmission was considerably higher (14.9%). CONCLUSION: V3 sequence analysis confirms the epidemiologic data which support the transmission of HIV-1 in children from a single source, and suggests the infection of a mother from her parenterally infected child. Furthermore, the genetic variability of HIV-1 V3 in the noncohort group was particularly divergent indicating the heterogeneity of the virus circulating in the former Soviet Union.

PIP: In 1988, an HIV-1 epidemic occurred in Elista, Kalmyk Republic, Russia, among 90 children in two hospitals after exposure to blood contaminated needles from an HIV infected infant. A few months later, a similar HIV-1 outbreak in children occurred in Rostov-on-Don, Russia, probably a result of transporting children from Elista to Rostov-on-Don hospitals. In Rostov-on-Don, it appears that seven HIV infected infants transmitted HIV to their mothers during breast feeding. Health workers collected blood samples from 22 HIV-1 infected subjects in Elista (n = 12) and Rostov-on-Don (n = 10 including 1 mother-child pair) and from 8 control subjects who became infected with HIV-1 via sexual (7) and parenteral (1) transmission from Minsk, Belarus. Researchers wanted to determine the extent of the diversity of proviral DNA encoding the V3 loop from different patients in the children cohort. They used nested polymerase chain reaction on DNA of primary peripheral blood mononuclear cells and then cloned and sequenced them to detail the HIV-1 V3 encoding region. The Elista and Rostov-on-Don samples shared common consensus sequences (127 nucleotide sequences) in the V3 region. The average mean interperson variation between the nucleotide sequences of HIV patients infected through sexual transmission from Minsk was 14.9%, which was much higher than those for Elista and Rostov HIV patients infected through parenteral transmission (5.9% and 6.6%, respectively). The major nucleotide sequence in the mother in the Rostov group, who was presumably infected with HIV by her HIV infected infant during breast feeding, matched that of her daughter. The mother had no history of blood transfusion or any other risk factors except breast feeding. These findings confirm that the Elista and Rostov groups shared a common HIV source. They also suggest that breast feeding was the route of HIV transmission for the mother. The genetic variability of HIV-1 V3 in the control group demonstrated the heterogeneity of HIV-1 in the former USSR.

Comparative humoral responses to HIV-1 p24gag and gp120env in subjects from East Africa and the UK.

We compared 1616 sera from HIV-1-infected subjects and matched HIV-negative local controls in Uganda, Kenya and the UK. Sera were screened for specific antibody to HIV-1 p24 Gag and gp120 Env proteins and for p24 antigenaemia. In contrast to the UK, the majority of African HIV-1-infected subjects maintained detectable anti-p24 antibodies. However, lower reactivity of anti-p24 was observed in African AIDS patients, compared with those with asymptomatic HIV-1 infection. This reduction in anti-p24 reactivity with more advanced clinical stage was less marked in African HIV-1 infection than in the UK. Correspondingly, p24 antigenaemia was more common in patients with AIDS from the UK than in African patients (65 versus 4%). Reductions in anti-gp120 reactivity were observed in African AIDS patients, compared with the asymptomatic group. However, median reactivity of anti-gp120 in UK patients remained unchanged in both asymptomatic and AIDS subjects. The differences in humoral response to p24 and gp120 between Africa and the UK are semi-quantitative rather than qualitative and could be explained by initial higher antibody response to HIV-1 in African subjects.

Decline of anti-p24 antibody precedes antigenaemia as correlate of prognosis in HIV-1 infection.

A cohort of 84 homosexual men presenting with persistent generalized lymphadenopathy (PGL) was studied between March 1982 and March 1987. A progression rate to AIDS of 5% per year was seen in those who were infected with HIV. Certain clinical features and routine laboratory investigations were significantly associated with an increased risk of disease progression, but had only limited predictive value. Two hundred and fifty-two serial sera from 57 patients were analysed for the p24 antigen (Abbott), the principal core protein of HIV and antibody against p24 by direct enzyme-linked immunosorbent assay (ELISA). Patients who remained well retained a high titre of anti-p24 antibody compared with those who progressed to AIDS-related complex (ARC) or AIDS. HIV antigen was detectable in 40% of AIDS patients 2 years before diagnosis, but antigenaemia was preceded by loss of anti-p24 antibody by up to 18 months and preceded AIDS by up to 40 months. ARC patients tended to be negative when tested for both alpha-p24 antibody and antigen, suggesting a transitional state. Analysis of the humoral response against gag proteins appears to correlate closely with clinical status and may be an earlier and more consistent way of predicting disease progression than p24 antigenaemia, or clinical and routine laboratory investigations.

Identification of an env G subtype and heterogeneity of HIV-1 strains in the Russian Federation and Belarus.

OBJECTIVE: To identify HIV-1 envelope sequence subtypes in infected individuals from the Russian Federation and Belarus. PATIENTS: A cohort of children infected after exposure to non-sterile needles during the 1988-1989 HIV-1 epidemic in southern Russia (n = 20) and HIV-1-seropositive individuals from Russia (n = 1) and Belarus (n = 7) infected via sexual transmission. METHODS: DNA samples derived from peripheral blood mononuclear cells were analysed for their HIV-1 genotypes by the heteroduplex mobility assay (HMA). The 1.3 kilobase-pair env gene fragments encoding a portion of gp120 were amplified by nested polymerase chain reaction, cloned and sequenced. The env sequences derived from these patients were aligned and phylogenetic neighbour-joining and maximum parsimony-derived trees generated. RESULTS: The env sequences derived from eight individuals infected in Russia and Belarus belong to subtype A (one), B (four), C (two), and D (one). Sequences derived from children, infected during parenteral manipulations in southern Russia, and one mother were closely related, but highly divergent, as a group, from all prototypic strains (genetic divergence, 17.2-22.9%). However, they clustered together with env sequences of the V1525 and LBV21-7 isolates from Gabon, recently described to be members of a new HIV-1 env subtype G. CONCLUSION: Extensive heterogeneity of HIV-1 subtypes was evident in the Russian Federation and Belarus. Our data also support the existence of an HIV-1 env genetic subtype G, and such isolates are now apparently present on both the African and European continents. These variants were identified through V3 peptide enzyme-linked immunosorbent assay screening and subsequent HMA analysis. The combination of these techniques represents a model for screening HIV variants within a large population.

Lack of correlation between V3-loop peptide enzyme immunoassay serologic subtyping and genetic sequencing.

OBJECTIVE: To compare the performance of V3-loop peptide enzyme immunoassay (PEIA) methodologies from four different laboratories for subtyping HIV-1, and to determine the causes for the lack of correlation between V3-loop PEIA serotyping and subtyping by sequencing. MATERIALS AND METHODS: Synthetic peptides derived from the amino-acid consensus sequences of the V3-loop of group M strains representing genetic subtypes A-F as well as reference strains were evaluated in PEIA by four different laboratories for their ability to accurately determine the subtype in a panel of 85 sera obtained from persons infected with known HIV-1 subtypes (28 subtype A, 34 subtype B, four subtype C, 10 subtype D, seven subtype F, one each of subtype H and G). Furthermore, the V3 loop of the corresponding virus was compared with the V3 loop of the peptides used in PEIA. RESULTS: The correlation between HIV-1 subtyping by sequencing and V3-loop PEIA from the different laboratories varied considerably for the different HIV-1 subtypes: subtype A (46-68%), B (38-85%), C (75-100%), D (29-50%), and F (17-57%). A 70% agreement between PEIA and sequencing subtypes was observed for samples with the concordant presence of the same octameric sequences in the V3 loop of the virus and the V3 loop of the peptide used in PEIA; however, only 42% of specimens with different V3-loop octameric viral and peptide sequences yielded concordant results in V3-loop serotyping and genetic subtyping. CONCLUSION: Our results indicate that V3-loop PEIA methodologies used in different laboratories correlate poorly with genetic subtyping, and that their accuracy to predict HIV-1 subtypes in sera of Belgian individuals infected with different HIV-1 subtypes (A, B, C, D, F, G and H) vary considerably. The poor correlation between serotyping and genetic subtyping was partly due to the simultaneous occurrence of subtype-specific octameric sequences at the tip of the V3 loop of viruses belonging to different genetic subtypes.

Immunization of human HIV-seronegative volunteers with recombinant p17/p24:Ty virus-like particles elicits HIV-1 p24-specific cellular and humoral immune responses.

OBJECTIVE: To evaluate the immune response to HIV-1 p24 generated in vivo by p17/p24:Ty virus-like particles (p17/p24:Ty-VLP) by examining the lymphoproliferative and antibody (Ab) responses to HIV-1 p24, as well as Gag-specific cytotoxic T lymphocytes (CTL), in HIV-seronegative volunteers immunized with hybrid p17/p24:Ty-VLP. DESIGN AND METHODS: Sixteen HIV-seronegative volunteers were immunized with p17/p24:Ty-VLP at two dose levels (100 or 500 micrograms) and monitored for the following 48 weeks for production of anti-p24 and anti-p17 Ab, in vitro lymphoproliferative responses to HIV-1 p24 and p17, and in vitro CTL responses to HIV-1 Gag. RESULTS: Twelve out of the 16 volunteers had significant p24-specific proliferative responses, with volunteers on the higher dose schedule exhibiting earlier proliferative responses than those on the lower dose schedule. Proliferative responses in both volunteer groups were similar in overall magnitude but appeared at different times during the immunization schedule. Anti-p24 Ab were detected in six out of the nine individuals in the lower dose group and in five out of the seven in the higher dose group. There was a good correlation between the presence of p24-specific Ab and the detection of lymphoproliferative responses to the p24 protein in peripheral blood mononuclear cells isolated from the same individuals. Anti-p17 Ab were detected in five volunteers. No Gag-specific CTL responses were detected. CONCLUSION: We conclude that hybrid HIV-1 p17/p24:Ty-VLP are capable of inducing both cellular and humoral immunity to HIV-1 Gag p17 and p24 components and are worthy of further study as a potential HIV immunotherapeutic.

Identification of HIV-1 subtype G from Uganda.

PIP: HIV-1 subtype G was identified by the authors in the former Soviet Union, and is related to a sequence VI525 from Gabon. All told, subtype G isolates have been identified in the former Soviet Union, Gabon, Zaire, and Nigeria. A subtype G gag sequence has also been reported in Taiwan. This paper reports the first HIV-1 subtype G to be isolated from Uganda. Uganda has the highest reported incidence of HIV-1 infection in Africa, with HIV-1 subtypes A, B, C, and D having already been isolated in the country. The authors isolated and sequenced HIV-1 genetic subtype G from a Ugandan female patient with CDC stage IV disease and a CD4 count of 20/cu. mm. Although recently immigrated to the UK from Uganda, there is epidemiologic evidence that the patient was infected with HIV-1 in Uganda. The V3 region of the isolate had 35 amino acids, with the common African tetrapeptide GPGQ at the tip of the loop. There were deletions relative to other G subtype sequences in the V4 and V5 env regions. The phylogenetic relationship of UGJW3 with other HIV-1 strains was inferred using nucleotide sequences and the PHYLIP package. The C2-V5 env region of the isolate was found to cluster with the subtype G isolates from the former Soviet Union and the Gabon. These findings confirm the need for further examination of the distribution of HIV-1 genetic subtypes and the exploration of their biologic significance.^ieng

Sequence analysis of an HIV type 1 env subtype E isolate from Thailand with discordant V3-loop serotyping.

PIP: HIV-1 genetic subtypes B and E are currently circulating in Thailand. Subtype E accounts for more than 90% of heterosexual transmission nationwide, while subtype B is transmitted mainly among IV drug users. This paper reports an HIV-1 Thai E isolate which yielded discordant results in serotyping and genotyping assays. An HIV-1-infected mother enrolled in Bangkok in a perinatal transmission study was identified independently as subtype B by V3 serotype by St. Mary's and HIV/AIDS Collaboration laboratories using different in-house assays. Her plasma was also screened for other HIV-1-specific immune responses, yielding a pattern of antibody reactivity similar to other subtype E sera. The genetic subtype of the isolate was identified by heteroduplex mobility assay (HMA) to further characterize it. Analysis results suggest that the woman was infected with HIV-1 subtype E. The env region encoding C2-V3 was subsequently sequenced to clarify the discordant results between the V3 serotype B and the genetic subtype E.^ieng

Serotyping HIV type 1 by antibody binding to the V3 loop: relationship to viral genotype. WHO Network for HIV Isolation and Characterization.

We have investigated whether peptides representing the HIV-1 principal neutralization domain (V3) can be used as antigens in antibody-binding assays to predict the genotypes of the subjects' virus. Serum samples collected from HIV-1-infected subjects from the four WHO-sponsored vaccine evaluation sites (Uganda, Rwanda, Thailand, and Brazil) were characterized by antibody binding to a panel of synthetic V3 peptides that were derived from the consensus sequences of the V3 region of the HIV-1 subgroups according to the env phylogenetic analysis (A-E). An indirect V3 peptide-binding assay was used for primary screening, and a V3 peptide antigen-limiting ELISA was then used as a secondary assay to discriminate cross-reactivity if the screening assay was equivocal. In general, V3 peptide serology could predict HIV-1 genotypes. In sera for which the genotype of the virus was known, peptide assays could predict the correct genotype in approximately 90% of cases for genotypes A, B, C, and E; Ugandan sera of genotype D were more broadly reactive. There was considerable serological cross-reactivity between some HIV-1 genotypes, in particular between A and C, and, to a lesser extent, B and D subtypes. Owing to polymorphism at the crown of the V3 loop, an additional B peptide (B') was required to type Brazilian B genotype sera. These simple assays may help facilitate the determination and distribution of HIV-1 genotypes circulating in populations.

Sequence analysis of the glycoprotein 120 coding region of a new human immunodeficiency virus type 1 subtype G strain from Russia.

PIP: This paper reports the sequencing of HIV-1 subtype G from the blood sample of a 39-year-old Russian male taken in November 1992. The heterosexual, White male and his wife living in Uzbekistan found in 1991 that they were infected with HIV-1. The man reported living in Mozambique during 1984-85 where he had no sexual relations with African residents, yet was admitted to the hospital on several occasions. However, in 1986-87, shortly after returning to the Soviet Union, he reported having a sexual relationship with a woman other than his wife. That woman and her husband were also found to be HIV-positive in 1991. The subject died in November 1993, and his wife in March 1995. His peripheral blood mononuclear cells were separated and kept at -70 degrees Celsius until used. When the blood sample was obtained, the man had a CD4 count of 10 cells/mcl, had lost more than 10% of his weight, and developed candidal esophagitis and chronic diarrhea associated with Cryptosporidia.^ieng

Genetic heterogeneity of HIV type 1 in Russia: identification of H variants and relationship with epidemiological data.

HIV-genetic subtypes were analyzed in 130 subjects from the Russian Federation, by the HMA technique. Six subtypes were identified in heterosexuals, including A, B, C, D, G, and H; however, homosexual men were infected predominantly with the B subtype (33 of 35). The subtype A isolates were found in population of intravenous drug users. HMA successfully identifies 128/130 DNA samples; the phylogenetic analysis of the V1/V5 gp120 encoding region derived from another two samples demonstrated that these isolate belong to subtype H.

An improved algorithm for determining HIV type 1 subtypes in a primary laboratory in Uganda.

A pilot study was undertaken with the objective of developing a simple, economical, and efficient algorithm through which to subtype HIV-1 in a large epidemiological cohort study in Uganda. A peptide enzyme immunoassay (PEIA) employing both V3 and gp41 regions and a heteroduplex mobility assay (HMA) were evaluated in comparison with DNA sequencing. Of 146 samples selected, 115 (79%) were successfully sequenced. Taking sequence data as the "gold standard," other assays were compared with these data. The HMA correctly identified 95 (83%) of the samples, and only 1 sample was wrongly identified. The V3 PEIA alone and in combination with gp41 peptides correctly identified 76 and 78% of the samples, respectively; however, the number of wrongly identified samples was four times less with the combination compared with V3 peptides alone (4 versus 16%). The sensitivity, specificity, and positive and negative predictive values for serotype A and D samples were greater for the combination than V3 peptides alone. We have described a new algorithm to segregate subtypes A and D. This algorithm uses the two peptide assays followed by HMA and then DNA sequencing for untypable samples, giving an accuracy of 95% at a cost of 37 and 21% for consumables compared with subtyping all the samples by HMA or DNA sequencing, respectively. This proposed approach is suitable for epidemiological studies in Uganda and other regions with a predominance of A and D subtypes.

An HIV type 1 epidemic among injecting drug users in the former Soviet Union caused by a homogeneous subtype A strain.

Epidemiological data have demonstrated rapid growth of HIV-1 infections among injecting drug users (IDUs) in the Ukraine and Russia, during 1996. Here we describe the results of genetic analysis of isolates derived from 12 HIV-1-infected IDUs in different sites of Russia and the Ukraine. The blood samples were taken within a 1- to 2-month period after the first HIV-1-positive test. The results of the heteroduplex mobility assay as well as gag/env phylogenetic analysis reveal that all sequences belong to gag/env genetic subtype A. Moreover, interpatient genetic distances between the nucleotide sequences encompassing the C2-V3, the V4-V5, and p17-encoding regions within this group were low (the average means were 0.9, 1.3, and 0.4%, respectively). These data show a marked homogeneity of HIV-1, probably spreading during primary infection. It is possible that the current epidemic of subtype A HIV-1 among IDUs in the former Soviet Union is caused by a point source exposure.

A sudden epidemic of HIV type 1 among injecting drug users in the former Soviet Union: identification of subtype A, subtype B, and novel gagA/envB recombinants.

The former Soviet Union republics have experienced an explosive human immunodeficiency virus type 1 (HIV-1) epidemic among injecting drug users (IDUs), consisting mainly of subtype A viruses originated from a point source (Bobkov et al.: AIDS Res Hum Retroviruses 1997;13:1195-1201). To determine whether new HIV-1 subtypes have entered the IDU population, 46 samples derived from IDUs in Russia (n = 39) and the Ukraine (n = 7) were genotyped by heteroduplex mobility assay (HMA). It was shown that 83% of IDU HIV-1 strains found in both countries belong to genetic subtype A. However, env subtype B was also found in 17% of cases. The sequence data showed a marked intrasubtype homogeneity of HIV-1 (the average means of interpatient genetic distance were 1.1 and 1.7% [in the gag gene] or 1.8 and 2.3% [in the env gene] for subtype A and subtype B, respectively), confirming the hypothesis of a point source of virus for each subtype variant. Moreover, recombinant gagA/envB variants originating from those two strains were also found in two samples collected in the Kaliningrad region of Russia. In conclusion, our results suggest that two strains of HIV-1 belonging to different genetic subtypes, A and B, as well as gagA/envB recombinants between genomes of these strains, are now circulating simultaneously among IDUs in the former Soviet Union.

Serotyping of HIV type 1 infections: definition, relationship to viral genetic subtypes, and assay evaluation. UNAIDS Network for HIV-1 Isolation and Characterization.

V3 serotyping refers to a system based on binding of antibody in patient sera to V3-loop peptides derived from HIV-1 env genetic subtypes. The V3x serotype represents reactivity of serum from an HIV-1-infected patient (regardless of viral genetic subtype), which reacts preferentially to a V3 peptide derived from the X subtype sequence. We have classified HIV-1 serotypes, determined the relationship between the HIV-1 V3 serotypes and viral genetic subtypes in a large study (n = 125), and evaluated the performance of three different V3 peptide-binding assays. Seven HIV-1 V3 serotypes were identified: A, B, B-Br, B-Th, C, D, and E. Serotypes B-Br and B-Th represent sera that react specifically to peptides derived from Brazilian B (B-Br, GWGR) and Thai B (B-Th, GPGQ) strains. The HIV-1 V3 B, C, and E serotypes correlated closely with their viral env genetic subtypes; 19-26 of 32 B sera (59-79%), 3-4 of 4 C sera (75-100%), and 19-22 of 23 E sera (83-96%) were identified as serotypes B, C, and E, respectively. In contrast, two major V3 serotypes were classified in A sera: A (14-18 of 36 [40-50%]) and C (12-19 of 36 [33-54%]). Similarly, two major V3 serotypes were classified in D sera: B (6-10 of 20 [30-50%]) and D (9-12 of 20 [45-60%]). Serotyping of subtype E sera showed the best concordance with genetic subtypes by all assays. Overall, HIV-1 V3 serotyping produced consistent results among three laboratories. However, HIV-1 V3 serotypes do not distinguish all HIV-1 genetic subtypes. The relative biological significance of the V3 serotypes remains to be elucidated.