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Biomedical subjects

R B Lal

Publications and source records attributed to R B Lal.

At least 91 records · Page 5Linked to original sources
PubMed1994

Interferon-gamma induces resistance in primary monocytes against human immunodeficiency virus type-1 infection.

Treatment of monocytes with interferon-gamma 1 day before, or at the time of infection with human immunodeficiency virus type-1 (HIV-1) induced complete resistance in monocytes against HIV-1 infection. There was no evidence of viral RNA, proviral DNA, p24 antigen, or reverse transcriptase activity through 2 weeks after inoculation. Ultrastructural examination of these cells showed no detectable virus particles. When interferon-gamma was added to monocytes 1 to 3 days post-infection, virus integration occurred, but the viral expression was either ablated (1 day post-infection) or significantly inhibited (3 days post-infection). Treatment of monocytes with interferon-gamma before or after infection with HIV-1 produced significantly higher levels of tumor necrosis factor-alpha and interleukin-8 than untreated or uninfected monocytes. These results suggest that altered regulation of cytokines may mediate antiviral activity of interferon-gamma in monocytes.

Cells, Cultured↗
PubMed1994

Modulation of HTLV-II-associated spontaneous lymphocyte proliferation by beta 2 integrin CD11a/CD18 involves interaction with its cognate ligand, CD54.

In vitro culture of lymphocytes from persons infected by human T-lymphocyte virus type II (HTLV-II) results in spontaneous proliferation in the absence of any exogenous stimuli. The present investigation examined the role of integrin molecules in spontaneous lymphocyte proliferation (SLP) in persons infected with HTLV-II (n = 18) and normal controls (n = 16). Phenotypic analysis of SLP cells on Day 8 demonstrated no change in the surface expression of CD29 (beta 1), CD49b,d,e, and f (alpha-chains) compared with cells from normal controls; however, there was an increase of CD29 expression on SLP cells on Day 8 (77.2 +/- 5.1%) compared with Day 0 (53.2 +/- 3.1%; P < 0.01). Furthermore, addition of extracellular matrix proteins, fibronectin, laminin, or collagen (beta 1 integrin ligands) did not alter either the proliferative responses or the adhesion clusters in either groups. Analysis of beta 2 integrins on SLP cells showed not only an increased cell surface density of both CD18 and CD11a but also differential expansion of CD8+ T-cells coexpressing CD18 (54.0 +/- 10.3%), CD11a (53.7 +/- 8.1%), and S6F1, an epitope of CD11a, (65.3 +/- 7.8%) on Day 8 compared with Day 0 (20.0 +/- 2.5%, 19.3 +/- 1.9%, and 38.0 +/- 7.0%, respectively). Monoclonal antibodies to CD18 and CD11a inhibited SLP by 55 +/- 6.3% in HTLV-II-infected persons in a dose-dependent manner. The inhibition of SLP by anti-beta 2 antibodies was not due to negative signaling, since these antibodies did not inhibit anti-CD3-stimulated proliferation of normal lymphocytes. Moreover, monoclonal antibodies to CD54, the ligand for CD11a, inhibited the SLP in the majority of HTLV-II-infected persons studied. Taken together, these data suggest that SLP by PBL from HTLV-II-infected individuals is mediated through increased expression of beta 2 integrins that can modulate cognate receptor/ligand interactions on the cell surface of autologous proliferating cells.

Antigens, CD↗
PubMed1994

Presence of human T lymphotropic virus types I and II in Ghana, west Africa.

Until recently, HTLV-I was considered to be an Old World virus and HTLV-II was thought to be endemic in the Americas. However, the presence of HTLV-II among Pygmies and other populations of Africa has raised doubts as to whether HTLV-II is primarily a New World virus. The large serosurveys conducted in the urban and rural areas of southern Ghana have identified a 1-2% prevalence for HTLV-I/II. To define the HTLV type, we have used a Western blot assay (HTLV-2.3 blot) that allows simultaneous confirmation and differentiation between HTLVs. Samples (n = 139) were chosen on the basis of previous reactivity with either an enzyme immune assay or r21e-spiked WB results. The WB 2.3 analysis of these specimens identified 55 (40%) to be HTLV positive, 70 (50%) to be HTLV indeterminant, and 14 (10%) to be HTLV negative for HTLV. HTLV seroindeterminant patterns ranged from both gag and env (14 were r21+, p24+, and/or p19+ [all were RIPA negative]) to gag only (21 were p24+/p19+, 16 were p19+, and 7 were p24+), and env only (8 were r21+ and 4 were rgp46+) reactivities. Of the 55 HTLV-positive specimens, 41 were typed as HTLV-I, 9 were HTLV-II, and 5 could not be typed (HTLV-I/II). Of the nine HTLV-II-positive specimens, three were from patients with Burkitt's lymphoma and six were from healthy individuals (two pregnant women) with no obvious risk factors for HTLV-II.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Viral↗
PubMed1994

Viral-specific humoral immune responses following transfusion-related transmission of human T cell lymphotropic virus type-I infection.

The immunoglobulin (Ig) isotypes of antibodies to specific proteins of the human T cell lymphotropic virus type I (HTLV-I) were determined by Western blot analysis of serial specimens from six individuals who experienced HTLV-I seroconversion following blood transfusion; five remained asymptomatic carriers, while one developed HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP) 32 weeks posttransfusion. Analysis of Ig isotypes demonstrated that while IgM was the most frequent early response to gag (p19, p24) and env (r21e) proteins within the first 3 months following transfusion, IgG and IgA responses could also be detected within this period. HTLV-I-specific antibody responses plateaued in all Ig isotypes, including IgM, within the next 4- to 6-month period following transfusion and persisted through the entire study period (> 4 years). Comparison of antibody profiles in Ig isotypes and IgG1 and IgG3 subclass among asymptomatic carriers and one individual who developed HAM/TSP demonstrated no evidence of isotypic prominence or IgG subclass restriction in either group. These results indicate the appearance of HTLV-I-specific IgM that persists even after the primary infection and suggest that such response does not appear to provide an early marker of seroconversion. Further, we found no evidence of isotypic prominence or restriction of the antibody response in recipients who remained asymptomatic compared to one who developed HAM/TSP.

Blotting, Western↗
PubMed1994

Sequence variation within the immunodominant epitope-coding region from the external glycoprotein of human T lymphotropic virus type II in isolates from Seminole Indians.

Western blot analysis of 16 serum specimens from Seminole Indians demonstrated that 14 reacted with the type-specific recombinant epitope (rgp46II+) of human T lymphotropic virus type II (HTLV-II), whereas the remaining 2 specimens did not (rgp46II-). Both rgp46II- specimens demonstrated presence of HTLV-II genome by polymerase chain reaction analysis. Culture of 1 of these specimens demonstrated presence of type C retrovirus particles by electron microscopy, and p24gag antigens were detectable in culture supernatant. Nucleotide sequence analysis of 557 bp in the env gene (position 5405-5961) from 2 each of the rgp46II- and rgp46II+ specimens demonstrated sequence conservation in the rgp46II epitope (K-55(162-205)). Thus, lack of immune reactivity to rgp46 is not due to sequence variation within this epitope. This observation suggests that immunodominant env epitopes may not be universally recognized. Therefore, specimens with p24gag and r21eenv reactivity in modified Western blot assays should be further tested by more sensitive techniques.

Amino Acid Sequence↗
PubMed1994

Differential immune responsiveness to the immunodominant epitopes of regulatory proteins (tax and rex) in human T cell lymphotropic virus type I-associated myelopathy.

Infection by human T cell lymphotropic virus type I (HTLV-I) is etiologically linked with HTLV-I-associated myelopathy (HAM) or adult T cell leukemia (ATL). To evaluate the contribution of the viral regulatory proteins tax and rex during the development of disease, antibody responses to these proteins were analyzed in patients with HAM (n = 28) or ATL (n = 48) and in asymptomatic carriers (n = 69). Epitope mapping analysis identified immunodominant epitopes towards the amino terminus (Tax8(106-125)) and at the carboxyl terminus (Tax22(316-335), Tax23(331-350), and Tax24(336-353)) of tax and the amino terminus (Rex1(1-20), Rex2(16-35), Rex4(46-65), and Rex6(76-95)) of rex. Analysis of antibody reactivity to these immunodominant epitopes demonstrated preferential reactivity to Tax8, Tax22, Tax23, and Tax24 (71%-93%) and to Rex4 and Rex6 (52%) in patients with HAM when compared with reactivities in ATL patients (4%-31% for tax and 19%-24% for rex) or asymptomatic carriers (27%-37% for tax and 7%-23% for rex). In contrast, antibody responses to the immunodominant epitopes of the env proteins of HTLV-I (MTA, Env1, Env5) were similar in all of three clinical groups. Thus, differential immune responsiveness to the immunodominant epitopes of tax and rex in patients with HAM may play a role in disease pathogenesis in HTLV-I-infected persons.

Amino Acid Sequence↗
PubMed1994

Immunologic cross-reactivity between structural proteins of human T-cell lymphotropic virus type I and the blood stage of Plasmodium falciparum.

To determine the serologic cross-reactivity between human T-cell lymphotropic virus type I (HTLV-I) and parasite antigens, we measured antibody responses against HTLV-I, Plasmodium falciparum, Plasmodium vivax, and Brugia malayi in serum specimens obtained from regions where malaria (n = 482) and filariasis (n = 101) are endemic. Analysis of immune reactivity to HTLV-I antigens showed that specimens from regions where malaria is endemic had significantly higher rates of enzyme immunoassay (EIA) reactivity (76 of 482 [15.8%] than those from regions where filariasis is endemic (0 of 101 [0%]). Western blot (immunoblot) analysis of the HTLV-I EIA-reactive specimens demonstrated predominant Gag reactivity (HTLV-Iind). Only two specimens each from Indonesia and Brazil and four specimens from Papua New Guinea had Env reactivity by radioimmunoprecipitation analysis. Furthermore, a positive correlation between HTLV-EIA and titers of antibody to the blood stage of P. falciparum (rs = 0.24, P < 0.005) was discerned; no correlation was observed between antibodies to the blood stage or the circumsporozoite protein of P. vivax and the circumsporozoite protein of P. falciparum. In addition, P. falciparum-infected erythrocyte lysate specifically abrogated binding of Gag-specific antibodies in HTLV-Iind specimens from regions where malaria is endemic without affecting binding in HTLV-I-seropositive specimens, suggesting that the immunologic cross-reactivity between HTLV Gag proteins and malaria parasites is restricted to the blood-stage antigens of plasmodia in specimens from regions where malaria is endemic. However, HTLV-seroindeterminate specimens from the United States did not demonstrate serologic cross-reactivity, suggesting that antigenic mimicry of HTLV proteins extends to other nonplasmodial antigens as well.

Animals↗
PubMed1994

Detection of antibodies to trans-activator protein (p40taxI) of human T-cell lymphotropic virus type I by a synthetic peptide-based assay.

Antibodies to human T-cell lymphotropic virus type I (HTLV-I) trans-activator protein (p40taxI) were determined in serum specimens from individuals infected with HTLV-I (n = 138) and HTLV-II (n = 19). Western blot (immunoblot) analysis using recombinant tax demonstrated the presence of anti-tax antibodies in 96% of patients (25 of 26) with HTLV-I-associated myelopathy, 43% of those (20 of 46) with adult T-cell leukemia, and 61% of asymptomatic HTLV-I blood donors (40 of 66); only one of the HTLV-II specimens reacted with the recombinant tax protein. Synthetic peptides (Tax8(106-125), Tax22(316-335), Tax-23(331-350), and Tax-24(336-353) representing the immunodominant epitopes of p40taxI detected anti-tax antibodies in 66 (48%), 50 (36%), 66 (48%), and 64 (46%) of 138 HTLV-I-positive specimens, respectively. An enzyme immunoassay using an equimolar ratio of these four peptides allowed sensitive detection of anti-tax antibodies in 96% of patients (25 of 26) with HTLV-1-associated myelopathy, 52% of adult T-cell leukemia patients (24 of 46), and 62% of asymptomatic HTLV-1-infected donors (41 of 66). The synthetic peptide-based cocktail assay was HTLV-I specific, since none of the HTLV-II-infected specimens reacted with these peptides. Interestingly, the corresponding regions from the HTLV-II tax protein, Tax8II(106-125), and Tax-22II(312-331) did not react with either HTLV-II or HTLV-I specimens. Thus, a synthetic peptide-based assay composed of immunodominant epitopes located towards the amino terminus and the C terminus of p40taxI provides a reliable and sensitive assay for the detection of anti-tax antibodies in seroepidemiologic studies.

Amino Acid Sequence↗
PubMed1994

Spontaneous lymphocyte proliferation in human T-cell lymphotropic virus type I (HTLV-I) and HTLV-II infection: T-cell subset responses and their relationships to the presence of provirus and viral antigen production.

Spontaneous lymphocyte proliferation (SLP) during in vitro culture of mononuclear cells (MCs) characterizes over half of asymptomatic individuals infected with human T-cell lymphotropic virus type I (HTLV-I) or HTLV-II. Both CD4 and CD8 T-cell subsets within MC cultures are activated during SLP, as judged by high-density CD25 (CD25bright) expression; it is unclear, however, whether both cell subsets can directly undergo SLP. In the present investigation, the SLP capacities of purified CD8 and CD4 cells were examined in subjects infected with HTLV-I (n = 19) or HTLV-II (n = 54) in relation to the SLP status of MCs from each subject. No increase in SLP was observed for CD8 or CD4 cells from SLP-negative (SLP-) HTLV-infected subjects, whereas robust SLP characterized CD8 cells from all SLP-positive (SLP+) individuals, regardless of HTLV type. In contrast, SLP+ CD4 cells characterized only 23% (7 of 31) of HTLV-II+ SLP+ individuals, whereas SLP+ CD4 cells characterized 100% of HTLV-I+ SLP+ individuals. In cocultures of HTLV-II+ SLP+ CD8 cells and autologous SLP- CD4 cells, sizable proportions of both CD8 cells and CD4 cells coexpressed CD25bright, suggesting that SLP- CD4 cells were activated in the presence of SLP+ CD8 cells. PCR analysis for tax sequences detected provirus in most CD4- and CD8-cell preparations from HTLV-seropositive individuals, regardless of type and the SLP status of cell subsets. To determine whether SLP was associated with activation of viral genes, levels of HTLV-I and HTLV-II core antigen (Ag) in supernatants were measured. Viral Ag production and SLP responses were significantly correlated for both CD4 and CD8 cells in both HTLV-I and HTLV-II infections. However, inhibition of CD8- or CD4-cell SLP by cyclosporin A or anti-Tac (anti-CD25) did not reduce Ag production, indicating that Ag production is not coupled to SLP. These findings show that CD4 cells from SLP+ HTLV-I+ and SLP+ HTLV-II+ individuals differ in SLP capacity, that the absence of SLP does not indicate a lack of infection, and that production of viral Ag is associated with, but not dependent on, SLP.

Base Sequence↗
PubMed1993

Infection with human T-lymphotropic viruses leads to constitutive expression of leukemia inhibitory factor and interleukin-6.

Leukemia inhibitory factor (LIF), similar to interleukin-6 (IL-6), is a glycoprotein growth factor and differentiation regulator that has pleiotropic activity in several cellular systems. Recent reports of constitutive IL-6 production from spontaneously proliferating cells from human T-cell leukemia virus (HTLV)-infected individuals led us to examine the expression of IL-6 and LIF during HTLV infection. In vitro infection of peripheral blood lymphocytes with HTLV-I was associated with production of both soluble LIF and IL-6 in conjunction with the increasing HTLV antigen concentration. Northern blot analysis of T-cell lines generated from individuals infected with HTLV-I (MT-2, HuT-102, FS, EG, SP) and HTLV-II (Mo-T, H2A, H2E) demonstrated a marked increase in constitutive expression of LIF and IL-6 transcripts, as compared with uninfected cell lines (HuT-78, Jurkat). The constitutive expression of LIF and IL-6 was independent of presence of IL-2 in the culture medium, as both IL-2-independent (MT-2, HuT-102, SP, Mo-T) and IL-2-dependent (FS, EG, H2A, H2E) cell lines expressed LIF and IL-6 transcripts. Furthermore, LIF and IL-6 RNA expression in an HTLV-I-infected cell line (MT-2) was enhanced by phorbol ester stimulation via mechanisms that appear to be dependent on the posttranscriptional regulatory controls. These results show that both LIF and IL-6 are produced by HTLV-I- and HTLV-II-infected cells, which could potentially alter the transcriptional regulation of HTLV gene expression by inducing certain early response genes.

Cell Line↗
PubMed1993

Constitutive expression and production of tumor necrosis factor-beta in T-cell lines infected with HTLV-I and HTLV-II.

A highly sensitive enzyme-linked immunoassay was developed to detect soluble levels of TNF-beta. Analysis of TNF-beta in culture supernatants from long-term T-cell lines infected with human T-cell lymphotropic virus (HTLV) types I and II demonstrated elevated levels of TNF-beta when compared with uninfected T-cell lines. Presence of interleukin-2 in the culture medium showed a synergistic effect upon TNF-beta production. Further, constitutive expression of TNF-beta mRNA was observed in most cell lines infected with HTLV-I or HTLV-II. These results demonstrate that infections with HTLV-I/II can alter production of TNF-beta, presumably via the transactivation of TNF-beta promoter by the tax protein of HTLV-I and HTLV-II.

Cell Line↗
PubMed1993

Idiopathic CD4+ T-lymphocytopenia--an analysis of five patients with unexplained opportunistic infections.

BACKGROUND: Although patients with idiopathic CD4+ T-lymphocytopenia and serious opportunistic infections have been described previously, the clinical and immunologic features of this condition have not been well defined. METHODS: We studied in detail five patients with idiopathic CD4+ T-lymphocytopenia. The studies included serologic testing, culture, and polymerase chain reaction for the human immunodeficiency virus (HIV) types 1 and 2, serologic testing for the human T-cell lymphotropic virus (HTLV) types I and II, lymphocyte phenotyping, immunoglobulin quantitation, and lymphocyte-transformation assays, as well as attempts to isolate a retroviral agent. The results were compared with those in HIV-infected persons matched for CD4+ T-cell counts and with those in normal controls. We also studied the spouses of patients and the blood donors for one patient. RESULTS: In these five patients, there was no evidence of either HIV or HTLV infection. All the patients had both low percentages and low counts of CD4+ T cells, with relative increases in percentages, but not counts, of CD8+ cells. Numbers of B cells and natural killer cells were generally normal. As compared with HIV-infected persons, our patients had lower percentages and counts of CD8+ cells and more lymphopenia. CD4+ counts were relatively stable over time. Instead of the high immunoglobulin levels seen in HIV infection, these patients had normal or slightly low levels of immunoglobulins. The lymphocyte-transformation response to mitogens and antigens was depressed. Results in spouses and blood donors were normal. CONCLUSIONS: Idiopathic CD4+ T-lymphocytopenia differs from HIV infection in its immunologic characteristics and in its apparent lack of progression over time. Nothing about the immunologic or viral-culture studies performed in these patients or about their family members or blood donors suggests that a transmissible agent causes this condition.

Adult↗
PubMed1993

Isotypic and IgG subclass restriction of the humoral immune responses to human T-lymphotropic virus type-I.

We have investigated the isotypic and IgG subclass profile of the antibody response to HTLV-I structural proteins (gag and env) in patients with HTLV-I-associated myelopathy (HAM; n = 20), adult T-cell leukemia (ATL; n = 15), and HTLV-I-positive asymptomatic carriers (ASY; n = 21). IgG, IgM, and IgA were the predominant antibody responses in all HTLV-I-infected individuals; minimal IgE response was detectable in the HAM and ATL groups. Among the IgG subclasses, IgG1 was the most predominant antibody detected in responses to HTLV-I antigens, followed by IgG3 and IgG2; IgG4 could not be detected in any patient group. Levels of both IgG1 and IgG3 were significantly higher in patients with HAM, when compared to ATL and ASY (P < 0.01 for both comparisons). In addition, Ig isotypes and IgG subclass antibody in patient sera reactive with purified viral proteins and several immunodominant epitopes, represented by synthetic peptides, Gag-1a102-117, Env-1(191-214), Env-5(242-257), and recombinant proteins, MTA-1(162-209) and r21e303-440, were examined to delineate specific epitopes responsible for inducing the host immune responses of each isotype and subclass to the structural proteins of HTLV-I. IgG, IgM, and IgA responses were directed against both the gag and env gene products. Among IgG subclasses, the IgG1 and IgG3 responses were directed against both the gag (p53, p24, p19, and Gag-1a) and env (recombinant MTA-1, r21e, and synthetic Env-1, Env-5) proteins; IgG2 responses were mainly restricted to gag proteins. The frequency profile of HTLV-I-specific antigen recognition in all four IgG subclasses were similar in all of the clinical groups. These results further define the fine specificity of anti-HTLV-I immune reaction for understanding the mechanism of pathogenesis in these individuals and suggest that factors other than the humoral immune responses may be associated with the clinical manifestation of the disease.

Amino Acid Sequence↗
PubMed1993

Isolation, characterization, and transmission of human T-lymphotropic virus types I and II in culture.

Highly sensitive coculture methods were developed both for isolation of human T-lymphotropic virus types I and II (HTLV-1 and HTLV-II) from infected individuals and for productive infection of lymphoid cells. Mitogen-activated peripheral blood mononuclear cells (PBMC) from 13 HTLV-I- and 20 HTLV-II-positive specimens were cocultured with an equal number of mitogen-activated PBMC from HTLV-seronegative individuals, and culture supernatants were tested for the presence of soluble p24gag antigens at weekly intervals for 4 weeks. Eleven of 13 (85%) HTLV-I and 14 of 20 (70%) HTLV-II cultures were positive for p24 antigens. None of the 17 HTLV-seroindeterminate or six HTLV-seronegative specimens were positive for the presence of p24 antigen. The isolation rates for HTLV-I and HTLV-II by an alternative whole-blood lysis procedure were comparable to those obtained by standard PBMC cultures. Furthermore, cocultivation of PHA-stimulated PBMC from healthy donors with lethally irradiated HTLV-I- and HTLV-II-infected cell lines (SP and Mo-T, respectively) resulted in productive viral infection, as reflected by the appearance of p24gag antigens concomitant with specific genomic amplification of HTLV proviral DNA after 3 weeks of cocultivation. Thus, the cocultivation technique provides a highly sensitive and specific procedure both for HTLV isolation and for infection of target cells.

Base Sequence↗
PubMed1993

Over expression of insulin-like growth factor receptor type-I in T-cell lines infected with human T-lymphotropic virus types-I and -II.

To determine if aberrant expression of tyrosine kinase growth factor receptors may be related to the cell transformation capabilities of human T-lymphotropic viruses (HTLVs), we examined the expression of the epidermal growth factor receptor (EGF-R), insulin receptor (INS-R), and insulin-like growth factor receptor type-I (IGFR-I) in cell lines infected with HTLV type I (MT-2, HuT-102) and HTLV type II (Mo-T). Levels of mRNA transcripts for IGFR-I were significantly higher in both MT-2, HuT-102 (HTLV-I) and Mo-T (HTLV-II) cell lines than in uninfected cell lines (HuT-78, Jurkat); no detectable levels of EGF-R or INS-R mRNA transcript were observed in HTLV-infected or uninfected cell lines. Southern blot analysis demonstrated that no amplification or rearrangement of the IGFR-I gene occurred in either the MT-2 or Mo-T cell line. Flow cytometry analysis demonstrated that while IGFR-I protein was constitutively expressed on the cell surface in both MT-2 and Mo-T cell lines, neither EGF-R nor INS-R proteins could be detected. Ligand binding studies with MT-2 and Mo-T cell lines demonstrating binding of 125I insulin-like growth factor type-1 (IGF-I) in a dose-dependent manner and this response could be inhibited by increasing concentrations of cold IGF-I. These data demonstrate that deregulated expression of functional IGFR-I, the regular component of the growth control machinery of normal cells, may contribute to cellular proliferation and eventual transformation in HTLV-I- and HTLV-II-infected cell lines.

Cell Transformation, Viral↗