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R B Lal

Publications and source records attributed to R B Lal.

At least 73 records · Page 4Linked to original sources
PubMed1996

Endemicity and phylogeny of the human T cell lymphotropic virus type II subtype A from the Kayapo Indians of Brazil: evidence for limited regional dissemination.

Long terminal repeat (LTR)-based restriction fragment length polymorphism (RFLP) analysis of human T cell lymphotropic virus type II (HTLV-II) from 17 seropositive Kayapo Indians from Brazil showed that all 17 samples contained a unique HTLV-IIa subtype (A-II). Additional RFLP screening demonstrated the presence of this subtype in two of three Brazilian blood donors and a Mexican prostitute and her child. In contrast, 129 samples from blood donors and intravenous drug users (IDUs) from the United States, two Pueblo Indian samples, five samples from Norwegian IDUs, and two samples from blood donors from Denmark were all found to be a different HTLV-IIa subtype (A-III). Phylogenetic analysis of two Kayapo and one Mexican LTR sequences showed that they cluster with a subtype A-II sequence from a Brazilian blood donor and with sequences from two prostitutes from Ghana and Cameroon. These results demonstrate that infection with the A-II subtype is endemic among the Kayapo Amerindians, has disseminated to non-Indian populations in Brazil, and is also present in Mexico. Furthermore, the A-II subtype does not appear to represent an origin for the HTLV-IIa infection in urban areas of the United States and Europe. This study provides evidence that HTLV-IIa may be a Paleo-Indian subtype as previously suggested for HTLV-IIb.

Base Sequence↗
PubMed1996

Sensitivity and specificity of a DNA polymerase chain reaction nonisotopic-based detection method for the confirmation of infection with human T-lymphotropic virus types I and II.

BACKGROUND: A convenient, standard format for the detection of polymerase chain reaction (PCR) amplicons would increase the use of PCR for the confirmation of infection with human T-lymphotropic virus types I and II (HTLV-I and HTLV-II). OBJECTIVES: To determine the sensitivity and specificity of an enzyme oligonucleotide assay (EOA) for the confirmation of infection with HTLV-I or HTLV-II. STUDY DESIGN: The sensitivity of the EOA was determined by examining 88 specimens representing diverse geographic-associated genotypes and clinical manifestations. The specificity was determined by testing 40 HTLV-seroindeterminate (PCR-negative) specimens. RESULTS: Of the 52 HTLV-I-positive specimens tested, 46 (88%) were confirmed positive for HTLV-I by the EOA; these included 25 of 30 (83%) specimens from asymptomatic carriers, 14 of 15 (93%) specimens from patients with HTLV-I-associated myelopathy, and all 7 specimens from patients with adult T-cell leukemia. Similarly, 33 of 36 (92%) HTLV-II-positive specimens were confirmed positive for HTLV-II. None of the specimens were wrongly classified. All specimens tested with distinct geographic-associated genotypes for HTLV-I and -II were detected by EOA. Analysis of seroindeterminate specimens, all of which were previously shown to be negative by nested PCR, showed that none of 40 were detected by either the HTLV-I or HTLV-II EOA. CONCLUSIONS: The overall sensitivity of the EOA detection for confirmation of HTLV-I and HTLV-II was 79 of 88 (90%) and the overall specificity was 100%. These findings demonstrate that the EOA provides a simple, standardized assay system for reliable confirmation and typing of HTLV infection.

Journal Article↗
PubMed1996

Delineation of immunodominant epitopes of human T-lymphotropic virus types I and II and their usefulness in developing serologic assays for detection of antibodies to HTLV-I and HTLV-II.

Several immunodominant B-cell epitopes have been mapped in the structural (gag, env) and regulatory (tax, rex) proteins of human T-lymphotropic virus type I (HTLV-I) and HTLV-II. Identification of these immunogenic epitopes not only has allowed the development of sensitive and specific serologic assays, but also has provided a tool to study correlates of host pathogenesis, viral transmission, and protection. The major immunogenic epitopes for HTLV-I are located at the C terminus of p19gag, the central and carboxyl terminus of gp46env, the central region of transmembrane envelope glycoprotein (p21e), and the carboxyl terminus of p40tax. Similarly, HTLV-II epitopes are located at the amino terminus and central region of gp46env, the central region of p21e, and the carboxyl terminus of p40tax. The transmembrane epitopes of HTLV-I and HTLV-II share extensive homologies and represent a highly cross-reactive region, while the remaining regions represent HTLV type-specific epitopes. The transmembrane epitope has been commonly used for detecting antibodies in both HTLV-I and HTLV-II-infected persons, whereas central envelope epitopes have been exploited for serologic differentiation between HTLV-I and HTLV-II. This detailed analysis at the epitope level has resulted in the development of highly sensitive and specific screening and confirmation assays for detection of antibodies to HTLVs.

Antibodies, Viral↗
PubMed1996

Forum on economic credentialing (E.C.).

What is economic credentialing? The California Medical Association defines economic credentialing as follows: "The use of economic criteria that do not apply to quality for granting or renewing medical staff pr ivileges?" The American Medical Association defines economic credentialing as " ... the use of economic criteria unrelated to quality of care or professional competency in determining an individual's qualification for initial or continuing hospital medical staff membership or pr-ivileges?" The Florida Medical Association defines economic credentialing as " ... any practice that denies access to hospitals based on eco- nomic criteria unrelated to the clinical qualifications or professional responsibilities of the physician." It also defines economic credentialing as " ... fiscal responsibility in practicing quality healthcare," and specifically notes that the governing body of a hospital has the right to discipline physicians (and presumably, exclude them) on the basis of resource utilization.

Journal Article↗
PubMed1995

In vivo cellular tropism of human T-lymphotropic virus type II is not restricted to CD8+ cells.

We have examined the in vivo and in vitro susceptibility of lymphocyte subpopulations to human T-lymphotropic virus type II (HTLV-II) to determine the cellular tropism for this virus. Monoclonal antibodies to T-cell subsets were used to separate highly purified CD4+ and CD8+ cells from peripheral blood lymphocytes of 35 individuals previously shown to be infected with HTLV-II. The purified T-cell subsets were analyzed for HTLV-II provirus (pol and tax gene sequences) by polymerase chain reaction (PCR) and cultured to determine virus expression by p24gag antigen detection. On the basis of PCR amplification in the pol and tax gene regions, both CD8+ subsets (89 to 91%) and CD4+ subsets (54 to 80%) from most infected subjects demonstrated HTLV-II provirus, irrespective of the viral genotype. Analysis of cultured lymphocytes demonstrated a higher spontaneous lymphocyte proliferation (17,986 +/- 4675 cpm) and p24gag antigen production (median 115 pg/ml; range 14-1360 pg/ml) in CD8+ cells compared to CD4+ cells (2333 +/- 826 cpm; p24gag antigen; 9 pg/ml; 2-250 pg/ml), suggesting a higher proviral load in CD8 cells. Limiting cell-dilution PCR analysis indicated that the CD8+ subset carried a higher HTLV-II provirus burden than the CD4+ subset. In vitro infection of purified CD4+ and CD8+ lymphocytes with irradiated HTLV-II cell lines resulted in productive infection of both subsets. Cell sorting and PCR analysis of lymphocyte subsets from 4 HTLV-II-infected subjects further demonstrated that in addition to CD4+ and CD8+ subsets, both CD45RO+ and CD45RO- and non-T-cells (CD14, CD16, and CD19) carried HTLV-II provirus. Taken together, these data suggest that HTLV-II possesses a broad tropism for peripheral blood mononuclear cells.

Antibodies, Monoclonal↗
PubMed1995

Molecular analysis of human T-cell lymphotropic virus type II from Wayuu Indians of Colombia demonstrates two subtypes of HTLV-IIb.

Studies of the genetic heterogeneity of human T-cell lymphotropic virus type II (HTLV-II) have revealed the presence of two genetic subtypes, termed HTLV-IIa and HTLV-IIb. The HTLV-IIb subtype encodes an immunodominant epitope present at the C-terminus of the extended Tax protein and, by using an LTR-based, restriction fragment-length polymorphism (RFLP) assay, can be further classified into IIb60-IIb5, with HTLV-IIb1 (Central Amerindian-like) and HTLV-IIb5 (North Amerindian-like) being characteristic subtypes for Native American Indians. To determine the antigenic and genetic heterogeneity among HTLV-II-infected South Amerindians, we used a Tax synthetic peptide immunoassay on serum, and RFLP and phylogenetic analysis on LTR sequences amplified from genomic DNA from four Wayuu Indians of Colombia. The Wayuu specimens displayed seroreactivity to the immunodominant epitope located in the extended Tax region, as predicted, and demonstrated genetic heterogeneity by the presence of both the IIB1 (Wyu1, Zuc31) and IIb5 (Wyu2, Zuc42) subtypes sequences within separate phylogroups represented by the Guaymi Indian (IIb1) and North Amerindian (IIb5) sequences, respectively. Sequence analysis showed that major LTR regulatory motifs and the cis-acting repressive elements in the LTR RNA secondary structure were relatively conserved in both Wayuu subtypes, but the predicted secondary structure of the rex response stem loop in the Wyu2 (IIb5) LTR sequence was 45 nucleotides (nt) and 95 nt longer than that observed in the Wyu1 (IIb1) and G12.1 (IIb1) LTR sequences, respectively. These results extend our knowledge of the genetic heterogeneity of HTLV-II in South Amerindians.

Amino Acid Sequence↗
PubMed1995

Genotypic characteristics of HTLV-II isolates from Amerindian and non-Indian populations.

The Amerindian human T-cell lymphotropic virus type II isolate HTLV-IIG12 has been demonstrated to be an HTLV-IIb with several unique features, including several restriction enzyme site changes, a distinctive pre-gag region, a stop codon within the pol gene, and an extended Tax protein. In this study, HTLV-II isolates from Amerindian and non-Indian populations were characterized by restriction enzyme site analysis to determine the prevalent HTLV-II subtype. In addition, DNA amplification by the polymerase chain reaction and Southern blot analyses were used to probe for the HTLV-IIG12 pre-gag region. Our results showed that of 13 Guaymi Indian isolates subtyped, all were HTLV-IIb, and that approximately one third of 17 isolates had the unique pre-gag region. While other HTLV-II-infected groups contained both HTLV-IIa and HTLV-IIb isolates, none of these isolates showed evidence of the distinctive HTLV-IIG12 pre-gag region. Lastly, DNA sequence analysis was used to determine the prevalence of the stop codon within the pol gene open reading frame. These analyses revealed that the occurrence of a stop codon within this sequence appeared to be characteristic of most HTLV-IIb subtypes. These results further our understanding of the genetic variations and evolution of the HTLV-II viruses within the endemically infected Amerindian populations, as well as U.S. intravenous drug users and other non-Indian populations.

Base Sequence↗
PubMed1995

Characterization of a HTLV-I-infected cell line derived from a patient with adult T-cell leukemia with stable co-expression of CD4 and CD8.

A long-term T-cell line, termed SP+, was developed from a human T-cell leukemia virus type I (HTLV-I)-infected patient with adult T-cell leukemia that is dependent on exogenous IL-2 for growth. The SP+ expresses a full complimentation of HTLV-I-specific viral proteins, and contains replication competent viral particles. Restriction enzyme digestion followed by Southern blot analysis demonstrated the presence of a single integrated proviral copy and limiting dilution analysis confirmed the clonality of the cell line. Interestingly, phenotypically, the SP+ cell line is CD2+, CD3+ and coexpresses CD4 and CD8, yet lacks TCR alpha beta and TCR tau delta expression. Further ontogenetic characterization of the SP+ cell line demonstrated the lack of thymic T-cell precursor markers, including absence of cell surface expression of CD1, intracellular thymic terminal deoxynucleotidyl transferase (TdT) enzyme, as well as message expression for V(D)J recombinase activating gene-1 (RAG-1). Furthermore, the SP+ cell did express the message for the CD3 delta chain. Taken together, these data suggest that the SP+ cell line resulted from HTLV-I infection of a mature CD4+/CDB+ lymphocyte. This cell line can be potentially useful as a model, both for regulation of cellular functions by HTLV-I and for immunologic functions of mature dual CD4/CD8 positive T-cells.

Antigens, Viral↗
PubMed1995

Mapping of linear epitopes of the regulatory proteins of human T cell lymphotropic virus (HTLV) type II: identification of an HTLV-IIb-restricted epitope.

Epitope mapping analyses using synthetic peptides representing the RexII and TaxII proteins identified predominant seroreactivity to the carboxyl terminus of the HTLV-IIG12 TaxII protein (G12Tax 22-G12Tax24, amino acids [aa] 312-356). Moderate reactivity to only 1 RexII peptide (G12Rex9, aa 121-140) was found, while all other RexII and TaxII peptides exhibited minimal reactivity. Peptide G12Tax24 (aa 337-356) corresponded to the extended portion of the TaxII protein characteristic of HTLV-IIb viruses and appeared to represent an HTLV-IIb-restricted epitope. This study showed that this peptide can be used in immunoassays as a quick, simple serologic tool for assessing the minimal number of HTLV-IIb viruses present within specific populations, especially when genomic DNA is not available.

Amino Acid Sequence↗
PubMed1995

Spontaneous proliferation of memory (CD45RO+) and naive (CD45RO-) subsets of CD4 cells and CD8 cells in human T lymphotropic virus (HTLV) infection: distinctive patterns for HTLV-I versus HTLV-II.

Spontaneous lymphocyte proliferation (SLP) in vitro is a characteristic feature of about 50% of individuals infected with HTLV-I or HTLV-II. Both CD4 cells and CD8 cells contribute to SLP in HTLV-I infection, whereas SLP in HTLV-II infection is usually restricted to CD8 cells. In this study, we asked if SLP was restricted to the memory (CD45RO+) cell subset of CD4 and CD8 cells in HTLV infection. Purified CD4 and CD8 cells were separated into CD45RO+ and CD45RO- populations by a modified panning technique, and spontaneous proliferation (SP) of the cell subsets was assessed. For all five HTLV-I-infected persons whose mononuclear cell cultures were SLP+, only CD45RO+ cells, but not CD45RO- cells, within CD4 and CD8 subsets showed SP. In contrast, five of six SLP+ HTLV-II+ individuals showed SP in both the CD45RO+ and the CD45RO- subsets of CD4 cells, and 10 of 12 SLP+ HTLV-II+ individuals showed SP of both the CD45RO+ and CD45RO- subsets of CD8 cells. Polymerase chain reaction studies showed that proviral genome was generally present in both CD45RO+ and CD45RO- subsets of CD4 and CD8 cells, regardless of HTLV type and SP activity. These findings show that SP of both CD4 and CD8 cells in HTLV-I infection is usually restricted to CD45RO+ memory cells, whereas in HTLV-II infection, both CD45RO+ memory and CD45RO- naive subsets of CD4 and CD8 cells may exhibit SP. It thus appears that HTLV-I infection and HTLV-II infection exhibit distinctive dysregulatory effects on memory and naive T cell subpopulations.

CD4-Positive T-Lymphocytes↗
PubMed1995

Infection with human T-lymphotropic virus types I and II results in alterations of cellular receptors, including the up-modulation of T-cell counterreceptors CD40, CD54, and CD80 (B7-1).

To examine the phenotypic alterations associated with human T-lymphotropic virus types I and II (HTLV-I and -II) infection, long-term cell lines (n = 12 HTLV-I cell lines; n = 11 HTLV-II cell lines; n = 6 virus-negative cell lines) were analyzed for the cell surface expression of various lineage markers (i.e., myeloid, progenitor, and leukocyte), integrin receptors, and receptor-counterreceptor (R-CR) pairs responsible for cellular activation. As expected, all cell lines expressed the markers characterizing the leukocyte lineage (CD43, CD44, and CD53). Of the progenitor-myeloid markers examined (CD9, CD13, CD33, CD34, and CD63), only the percent expression of CD9 was significantly increased on HTLV-I and -II-infected cell lines as compared with that on virus-negative cell lines. Analysis of the beta 1 integrin subfamily (CD29, CD49b, CD49d, CD49e, and CD49f) showed no significant change, except that CD49e was significantly decreased on the HTLV-infected cell lines. For the beta 2 integrin subfamily, the cell surface density was increased for CD18 and CD11a, while the CD11c molecule was expressed exclusively on the HTLV-I- and HTLV-II-infected cell lines. Analysis of several R-CR pairs (CD2-CD58, CD45RO-CD22, CD5-CD72, CD11a-CD54, gp39-CD40, and CD28-CD80) demonstrated that comparable levels of expression of the Rs (CD2, CD45RO, CD5, and CD28) and of some of the CRs (CD58, CD22, and CD72) were in all cell lines; however, CD54, CD40, and CD80 were expressed constitutively on the HTLV-I- and HTLV-II-infected cell lines. Functionally, the expression of these R-CR pairs did not appear to affect the autologous proliferation since monoclonal antibodies to these R-CR pairs were not able to inhibit proliferation of the infected cell lines. Taken together, our results indicate that HTLV-I and -II can modulate the expression of several T-cell activation molecules and CRs normally expressed on alternate cell types.

Antigens, CD↗
PubMed1995

Enhanced specificity of truncated transmembrane protein for serologic confirmation of human T-cell lymphotropic virus type 1 (HTLV-1) and HTLV-2 infections by western blot (immunoblot) assay containing recombinant envelope glycoproteins.

Immunoassays based on the highly immunogenic transmembrane protein of human T-cell lymphotropic virus type 1 (HTLV-1) (protein 21c) are capable of detecting antibodies in all individuals infected with HTLV-1 and HTLV-2. However, because of antigenic mimicry with other cellular and viral proteins, such assays also have a large proportion of false-positive reactions. We have recently identified an immunodominant epitope, designated GD21-I located within amino acids 361 to 404 of the transmembrane protein, that appears to eliminate such false positivity. This recombinant GD21-I protein was used in conjunction with additional recombinant HTLV type-specific proteins and a whole virus lysate to develop a modified Western blot (immunoblot) assay (HTLV WB 2.4). The sensitivity and specificity of this assay were evaluated with 352 specimens whose infection status was determined by PCR assay for the presence or absence of HTLV-1/2 proviral sequences. All HTLV-1-positive (n = 102) and HTLV-2-positive (n = 107) specimens reacted with GD21-1 in the HTLV WB 2.4 assay, yielding a test sensitivity of 100%. Furthermore, all specimens derived from individuals infected with different viral subtypes of HTLV-1 (Cosmopolitan, Japanese, and Melanesian) and HTLV-2 (IIa0, a3, a4, IIb1, b4, and b5) reacted with GD21-I in the HTLV WB 2.4 assay. More importantly, HTLV WB 2.4 analysis of 81 PCR-negative specimens, all of which reacted to recombinant protein 21e in the presence or absence of p24 and p19 reactivity in the standard WB assay, showed that only two specimens retained reactivity to GD21-I, yielding an improved test specificity for the transmembrane protein of 97.5%. None of 41 specimens with gag reactivity only or 21 HTLV-negative specimens demonstrated reactivity to GD21-I. In an analysis of additional specimens (n = 169) from different geographic areas for which PCR results were not available, a substantial increase in the specificity of GD21-I detection was demonstrated, with no effect on the sensitivity of GD21-I detection among specimens from seropositive donors. Thus, the highly sensitive, GD21-I-based HTLV WB 2.4 assay eliminates the majority of false-positive transmembrane results, thereby increasing the specificity for serologic confirmation of HTLV-1 and HTLV-2 infections.

Amino Acid Sequence↗
PubMed1995

Human T-lymphotropic virus type 1 peptides in chimeric and multivalent constructs with promiscuous T-cell epitopes enhance immunogenicity and overcome genetic restriction.

Conventional strategies of viral peptide immunizations often elicit low-affinity antibody responses and have limited ability to elicit immune responses in outbred animals of diverse major histocompatibility (MHC) haplotypes. This genetically restricted T-cell-stimulatory activity of peptides is a serious obstacle to vaccine design. However, the use of promiscuous T-cell epitopes may circumvent this problem. Promiscuous T-cell epitopes from tetanus toxin (amino acids [aa] 580 to 599) and the measles virus F protein (aa 288 to 302) that bind to several isotypic and allotypic forms of human MHC class II molecules have been identified and have been used in highly immunogenic constructs to overcome haplotype-restricted immune responses. Chimeric and beta-template peptide constructs incorporating known human T-lymphotropic virus type 1 (HTLV-1) B- and T-cell epitopes from the surface envelope protein gp46 (SP2 [aa 86 to 107] and SP4a [aa 190 to 209]) and promiscuous T-cell peptides were synthesized, and their immunogenicities were evaluated in both rabbits and mouse strains of divergent haplotypes (C3H/HeJ [H-2k], C57BL/6 [H-2b], and BALB/c [H-2d]). In addition, peptide preparations were structurally characterized by analytical high-performance liquid chromatography, mass spectrometry, and circular dichroism. In contrast to their linear forms, the chimeric constructs of both the SP2 and SP4a epitopes displayed alpha-helical secondary structures. Immunogenicity of the peptide constructs was evaluated by direct and competitive enzyme-linked immunosorbant assay (ELISA), as well as by radioimmunoprecipitation, syncytium inhibition, and antigen-induced lymphocyte proliferation assays. Immunization with the SP4a peptide without conjugation to a carrier protein produced antibodies specific for SP4a in two mouse strains (C3H/HeJ and C57BL/6). However, BALB/c mice failed to respond to the peptide, indicating that the T-cell epitope of the SP4a sequence is MHC restricted. In contrast, the chimeric constructs MVF-SP2 and SP4a-measles virus F protein were highly immunogenic, producing elevated ELISA titers after only two immunizations. Elicited antibodies recognized native forms of gp46 in ELISAs and radioimmunoprecipitation assays, as well as inhibited HTLV-1-mediated syncytium formation. In addition, chimeric constructs were effective at induction of lymphocyte proliferation to the T-cell epitope, SP4a, in each strain of immunized mice. Our data demonstrate that the antibody response to retroviral peptides is enhanced by promiscuous peptide constructs, in part because of the ability of such constructs to promote appropriate secondary structural forms of viral epitopes. In addition, these constructs promote virus-specific helper T-cell responses, thereby overcoming genetically restricted immune responses to the synthetic peptides.

Amino Acid Sequence↗
PubMed1995

Selective infection of human T-lymphotropic virus type 1 (HTLV-1)-infected cells by chimeric human immunodeficiency viruses containing HTLV-1 tax response elements in the long terminal repeat.

Previous studies have suggested that the human immunodeficiency virus long terminal repeat (HIV LTR) enhancer/promoter sequences contribute to the replication ability of HIV in different T-cell lines; mutation of these sequences can alter HIV tropism. We have utilized site-specific mutagenesis to generate variants of HIV that exhibit specific tropism for human T-lymphotropic virus type 1 (HTLV-1) Tax-expressing CD4+ T cells. The wild-type HIV LTR NF-kappa B and Sp1 sites in an infectious molecular clone of HIV type 1 were replaced with sequences derived from the 21-bp Tax response elements (TRE) from the HTLV-1 LTR to generate TRE-containing chimeric HIVs (TRE-HIVs). The TRE-HIVs exhibit selective replication and cell killing in HTLV-infected human CD4+ T cells, but not in HTLV-negative T cells. Transient transfections suggested that Tax-TRE interactions could account for the observed replication specificity. The TRE-containing HIV LTRs were synergistically activated by the HIV Tat and HTLV-1 Tax transactivators. These results demonstrate that it is possible to specifically target HIV replication and cytotoxicity to HTLV-1+, CD4+ human T cells, on the basis of Tax-TRE interactions, and provide a model for the development of specific, cytotoxic, retroviral gene therapy vectors for HTLV-1-infected cells based on alterations of the LTR transcriptional regulatory elements. They also suggest that HIV Tat can cooperate with heterologous transcriptional activators, such as Tax, which act through upstream binding sites without directly binding to DNA.

Base Sequence↗
PubMed1995

Phylogenetic relationship and geographic distribution of multiple human T-cell lymphotropic virus type II subtypes.

The current env-based subtyping of human T-cell lymphotropic virus type II (HTLV-II) identifies only two heterogenetic groups, HTLV-IIa and HTLV-IIb. To better understand the genetic diversity and phylogeny of HTLV-II, we examined the most divergent genomic region of HTLV-II, the long terminal repeat, by using restriction fragment length polymorphism (RFLP) and sequence analysis. Long terminal repeat sequences were amplified from peripheral blood mononuclear cells by PCR and digested with seven restriction endonucleases that differentiated HTLV-II into five HTLV-IIa (IIa0 to IIa4) and six HTLV-IIb (IIb0 to IIb5) restriction types, with HTLV-IIa0 and HTLV-IIb0 being prototypes for the MoT and NRA isolates, respectively. We examined 169 HTLV-II-infected samples, including 123 from blood donors and intravenous drug users (IDU) from the Americas, 16 from IDU from Europe, and 30 from Amerindians. Of the 169 samples, 109 (64.5%) were categorized as HTLV-IIa and 60 (35.5%) were categorized as HTLV-IIb. The predominant restriction types seen among the U.S. blood donors and U.S. IDU were IIa0 (68.7%) and IIb4 (10.4%). Four Spanish and seven Italian samples were IIb4, while five Norwegian samples were IIa2. Twelve Guaymi and all ten Seminole samples were single restriction types (IIb1 and IIb5, respectively), whereas the two Navajo and six Pueblo samples had a mixture of restriction types IIa0, IIa4, and IIb5. Of the HTLV-IIb restriction types observed in the U.S. non-Indians, 42.8% appear to have originated from the North Amerindian (IIb5), while 57.2% were similar to the European IIb4 restriction type. Sequences of 15 selected HTLV-II samples were determined and phylogenetically compared with 7 previously published HTLV-II LTR sequences. The derived topologies revealed three HTLV-IIa phylogroups (A-I to A-III) and four HTLV-IIb phylogroups (B-I to B-IV). Furthermore, the HTLV-IIa phylogroups appear to have evolved from the HTLV-IIb phylogroups. In the HTLV-IIa cluster, a Navajo (A-I) and a Brazilian (A-II) sequence formed separate phylogroups, while the remaining IIa sequences formed a single phylogroup (A-III). The four HTLV-IIb phylogroups were represented predominantly by a New York IDU (B-I), European IDU (B-II), North Amerindian and NRA (B-III), and Central Guaymi Indian (B-IV) sequence(s). Comparison of the phylogenetic data with the RFLP results revealed that results of the two methods correlated completely, demonstrating the ability of the RFLP method to predict the phylogroup of HTLV-II-infected samples accurately and quickly. GENBANK/U10258

Base Sequence↗
PubMed1995

Association between maternal antibodies to the external envelope glycoprotein and vertical transmission of human T-lymphotropic virus type I. Maternal anti-env antibodies correlate with protection in non-breast-fed children.

Vertical transmission of human T-lymphotropic virus type I (HTLV-I) depends primarily on breast-feeding; substitution of bottle-feeding has reduced the transmission rate from 20% in breast-fed children to 3% among bottle-fed. To determine the correlates of transmission for long breast-feeding (> or = 6 mo), short breast-feeding (< 6 mo), and bottle-feeding mothers, the antibody titers of transmitter (T) mothers and non-transmitter (nT) mothers were analyzed by using synthetic and recombinant epitopes representing the immunodominant epitopes of gag (Gag1a, r24), env (Env1/5, MTA1, RE3), and tax (Tax8/22-24) proteins. Seroreactivity to gag and tax epitopes was not significantly different except for anti-r24 antibody titer, which was significantly higher among T-mothers (geometric mean 134) when compared with nT-mothers (62) in the long-feeding group (P < 0.001). Profiles of antibody titers against env epitopes were different. Within the long-feeding group, Env1/5, MTA1, and RE3 titers were significantly higher among T-mothers (258, 1,476, and 738, respectively) when compared with nT-mothers (106, 279, and 320, respectively) (P < 0.01 for all three epitopes). In contrast, within the bottle-feeding group, antibody titers to Env1/5 (269) and RE3 (418) among nT-mothers were significantly higher than those among T-mothers (80 and 113, respectively) (P < 0.01). These data confirm that high-titered anti-HTLV-I antibodies in the long-feeding group correlate with milk-borne transmission of HTLV-I and, more importantly, imply that maternal anti-env antibodies may reduce the risk of non-milkborne infection.

Amino Acid Sequence↗
PubMed1995

Evaluation of immunodominant epitopes of human T-lymphotropic virus type 1 (HTLV-I) using synthetic peptides.

Human T-lymphotropic virus type 1 (HTLV-I) causes adult T-cell leukemia/lymphoma (ATLL) and has been associated with a variety of immunologically-mediated diseases. Recently, the immunodominant epitopes of HTLV-I have begun to be defined through the utilization of synthetic peptides and recombinant proteins. Strategies to define the conformational features of immunogenic peptides and design chimeric and multivalent constructs that mimic native viral proteins have provided the opportunity to create an effective synthetic vaccine against HTLV-I infection. An ideal peptide vaccine to be universally immunogenic must incorporate rationally designed antigenic determinants that accurately mimic the corresponding structural architecture found in native proteins and elicit relevant components of the immune system. We have recently designed and tested chimeric and beta-sheet template constructs containing HTLV-I immunodominant peptide motifs that elicit neutralizing antibody responses and overcome genetically restricted immune responses. To further illustrate putative vaccine candidates, HTLV-I env and tax proteins were analyzed using various computer-predicted correlates of protein antigenicity, secondary structural predictions, and major histocompatibility complex class I binding motifs. These approaches provide the opportunity to design synthetic peptide vaccines against HTLV-I infection that are based on structurally defined criteria, as well as test the influence of glycosylation on peptide conformation and immunogenicity.

Amino Acid Sequence↗