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1.
J Clin Microbiol ; 37(5): 1269-73, 1999 May.
Article in English | MEDLINE | ID: mdl-10203469

ABSTRACT

A rapid procedure for the diagnosis of malaria infections directly from dried blood spots by PCR amplification was evaluated with samples from 52 patients. Plasmodium infections were identified with a genus-specific primer set, and species differentiation between Plasmodium falciparum and Plasmodium vivax was analyzed by multiplex PCR. The PCR test with any of the three primer sets was able to detect as few as four parasites per microliter by gel electrophoresis or by nonisotopic paper hybridization chromatography. The diagnoses obtained by PCR correlated closely with those obtained by Giemsa staining except for two samples observed to have mixed P. falciparum-P. vivax infections. These were initially missed by microscopic analysis. In comparison with antigen-capture assays for P. falciparum, the PCR assays were able to detect three infections that were missed by the ParaSight-F test. The PCR test was negative for nine ParaSight-F-positive samples and one ICT Malaria Pf-positive sample, and these were confirmed to be false-positive results. The PCR thus gave no false-negative or false-positive results. Patients undergoing antimalarial therapy were also monitored by the PCR assay. Four of seven patients who were PCR positive for P. vivax at the time of discharge were later readmitted to the hospital with a recurrence of P. vivax infection. We would like to propose that PCR is a sensitive and easy method that can serve as a useful addition to microscopy for the diagnosis and the clinical monitoring of treatment of malaria.


Subject(s)
Malaria/diagnosis , Plasmodium/isolation & purification , Polymerase Chain Reaction , Animals , Humans , Sensitivity and Specificity
2.
Antisense Nucleic Acid Drug Dev ; 8(2): 95-101, 1998 Apr.
Article in English | MEDLINE | ID: mdl-9593047

ABSTRACT

Human papillomavirus (HPV) type 16 E6 and E7 inactivate the tumor suppressors p53 and pRB, respectively. Both viral oncoproteins play important roles in maintaining the transformed phenotype of cells. In this study, we examine the effects of antisense oligodeoxynucleotides with polarity and anomeric center reversal (alpha/beta-ODNs). ODNs of the general structure 5'alphaN3'3'NNN5'5'alphaN3'3'NNNN5'5'alphaN3+ ++'3'N5' were synthesized using phosphoramidite DNA chemistry. These alpha/beta-ODNs were complementary in sequence to regions flanking the start codons of HPV type 16 E6 and E7 genes. The anti-HPV type 16 alpha/beta-ODNs were able to form stable duplexes with their complementary RNA, which then serve as substrates for RNase H hydrolysis. Anti-HPV type 16 alpha/beta-ODNs also specifically inhibited the growth of two cervical carcinoma cell lines, CaSki and SiHa, both of which harbor HPV type 16 DNA. A decrease in E7 protein expression was also observed. Injection of nude mice with SiHa cells induces tumors. Treatment of these tumor-bearing mice with anti-HPV type 16 alpha/beta-ODNs led to substantially smaller tumors. These results show that alpha/beta-ODNs can exert antisense activities both in vitro and in vivo on the E6 and E7 genes of HPV type 16.


Subject(s)
Oligonucleotides, Antisense/pharmacology , Oncogene Proteins, Viral/genetics , Papillomaviridae/genetics , Repressor Proteins , Animals , Carcinoma/pathology , Carcinoma/therapy , Cell Division/drug effects , Codon/genetics , DNA, Complementary/genetics , DNA, Viral/genetics , Female , Gene Expression Regulation, Viral/drug effects , Genes, Viral , Humans , Mice , Mice, Inbred BALB C , Mice, Nude , Neoplasm Transplantation , Nucleic Acid Hybridization , Oligonucleotides, Antisense/chemistry , Oligonucleotides, Antisense/therapeutic use , Oncogene Proteins, Viral/biosynthesis , Oncogenes , Papillomavirus E7 Proteins , Papillomavirus Infections/pathology , RNA, Viral/genetics , RNA, Viral/metabolism , Ribonuclease H/metabolism , Structure-Activity Relationship , Tumor Cells, Cultured , Tumor Virus Infections/pathology , Uterine Cervical Neoplasms/pathology , Viral Structural Proteins/genetics
3.
Gene ; 200(1-2): 91-8, 1997 Oct 24.
Article in English | MEDLINE | ID: mdl-9373142

ABSTRACT

The murine malaria parasite Plasmodium berghei contains a plastid-like extrachromosomal genome. This genome is 30.7 kb in size and is transcriptionally active as shown by RT-PCR. DNA sequence analysis of the genome reveals 69.9-95.5% homology to sequences of the 35-kb extrachromosomal circle found in the human malaria species Plasmodium falciparum. Homologous sequences include regions of genes for the ssu-rRNA, lsu-rRNA, rpo B and clusters of t-RNAs. Sequence variation between the two Plasmodium species exists in the non-coding interspacing regions. A physical map has been constructed for the P. berghei circle, indicating the EcoRI and HindIII restriction sites as well as the arrangement of the rRNA, rpo B and tRNA genes. Arrangement of these genes is similar to that found on the P. falciparum 35-kb circle. The P. berghei circular element is distinct from the mitochondrial 6-kb DNA of both the murine and the human Plasmodium species. Preliminary results indicate that the circle may be a useful target for drug therapy.


Subject(s)
DNA, Protozoan/chemistry , Plasmodium berghei/genetics , Animals , Base Sequence , DNA, Mitochondrial/chemistry , DNA, Mitochondrial/genetics , DNA, Protozoan/genetics , DNA, Protozoan/ultrastructure , Genetic Variation , Humans , Mice , Plasmodium falciparum/genetics , Plastids/ultrastructure , Polymerase Chain Reaction , RNA, Protozoan/biosynthesis , Restriction Mapping , Sequence Homology, Nucleic Acid
4.
Acta Trop ; 68(1): 105-14, 1997 Oct 14.
Article in English | MEDLINE | ID: mdl-9352006

ABSTRACT

The Plasmodium parasite possesses two extrachromosomal genomes; the mitochondrial genetic element and the extrachromosomal plastid-like DNA. The latter has only been fully described for one culture strain of P. falciparum. In this study, a rapid procedure for amplifying plastid DNA from dried blood spots of blood infected with different malaria species was developed. PCR amplification of a 595 bp fragment within the plastid-like large subunit ribosomal-RNA (LSU-rRNA) gene was achieved using primers derived from the P. falciparum sequence. The PCR product was observed in all Plasmodium species examined. Sequence analysis of amplified products homologous to an LSU-rRNA fragment of the plastid-like extrachromosomal circle revealed extensive conservation between Plasmodium species including P. falciparum, P. vivax, P. malariae and P. berghei.


Subject(s)
DNA, Protozoan/blood , Plasmodium/genetics , Polymerase Chain Reaction , Animals , Base Sequence , DNA, Protozoan/chemistry , Humans , Molecular Sequence Data , RNA, Ribosomal/chemistry , Sequence Alignment
5.
J Med Virol ; 50(1): 93-6, 1996 Sep.
Article in English | MEDLINE | ID: mdl-8890046

ABSTRACT

The BMRF1 protein is an Epstein-Barr virus (EBV) DNA polymerase accessory protein that forms part of the early antigen diffuse (EA-D) component. An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of IgA antibody to the BMRF1 protein of EBV in saliva and serum samples. The assay was shown to be specific for nasopharyngeal carcinoma (NPC) patients and, when used with saliva alone, to have a sensitivity comparable to an existing indirect immunoperoxidase assay for early antigens. The sensitivity of the assay could be significantly enhanced to 86% by the use of paired saliva and serum samples.


Subject(s)
Antibodies, Viral/blood , Antigens, Viral/immunology , Enzyme-Linked Immunosorbent Assay , Herpesviridae Infections/virology , Herpesvirus 4, Human/isolation & purification , Nasopharyngeal Neoplasms/virology , Saliva/virology , Tumor Virus Infections/virology , Antibodies, Viral/immunology , Herpesviridae Infections/blood , Herpesviridae Infections/immunology , Herpesvirus 4, Human/immunology , Humans , Immunoglobulin A/blood , Immunoglobulin A/immunology , Nasopharyngeal Neoplasms/blood , Nasopharyngeal Neoplasms/immunology , Saliva/immunology , Tumor Virus Infections/blood , Tumor Virus Infections/immunology
6.
Parasitol Res ; 82(2): 130-35, 1996.
Article in English | MEDLINE | ID: mdl-8825207

ABSTRACT

On the basis of immunological cross-reactivity, we identified a 43-kDa Plasmodium berghei antigen with homology to the exp-1 antigen from P. falciparium. The P. berghei antigen was recognized by an antibody directed against an epitope on the C-terminus of the P. falciparum exp-1 protein. This antigen is localized on the surface of the parasite and shares peptide sequence homology with the P. chabudi antigen Ag3008. To investigate further the role of the P. berghei antigen, we designed antisense phosphorothioate oligodeoxynucleotides (PS oligos) complementary to sequences of the exp-1 mRNA from P. falciparum. The PS oligos were capable of inhibiting the development of P. falciparum in vitro by 47%. In vivo, experiments in mice showed that the same PS oligos had the potential to extend the life span of mice infected with P. berghei by a factor of 2-4. The immunological cross-reactivity and the antisense inhibition of P. berghei parasite development in vivo indicate that this antigen may be a homologue of exp-1 from P. falciparum that has functional importance for parasite survival.


Subject(s)
Antigens, Protozoan/immunology , Plasmodium berghei/immunology , Plasmodium chabaudi/immunology , Plasmodium falciparum/immunology , Amino Acid Sequence , Animals , Antimalarials/pharmacology , Base Sequence , DNA, Protozoan/genetics , Malaria, Falciparum/drug therapy , Malaria, Falciparum/immunology , Malaria, Falciparum/pathology , Mice , Molecular Sequence Data , Oligonucleotides/pharmacology , Peptides/chemistry , Plasmodium falciparum/drug effects
7.
Cancer Res ; 55(20): 4599-605, 1995 Oct 15.
Article in English | MEDLINE | ID: mdl-7553636

ABSTRACT

Human cervical cancers are often associated with human papillomavirus (HPV). In HPV-positive cervical cancers, the oncoproteins E6 and E7 are consistently expressed. In this study, the effects of antisense inhibition of both proteins were examined. Phosphorothioate oligonucleotides (ODNs) AE6 and AE7 complementary to regions flanking the start codons of HPV16 E6 and E7 genes, respectively, were synthesized. These anti-HPV ODNs inhibited the growth of cervical cell lines CaSki and SiHa, which harbor HPV16 but had little effect on cells that do not. Both ODNs also affected the ability of CaSki cells to form colonies in soft agar. In nude mice, treatment with either AE6, AE7, or a mixture of both led to substantially smaller tumors. AE7 was observed to inhibit E7 synthesis. The AE6 ODN probably exerts its effect by suppressing the expression of E6 as well as E7. Cell cultures and tumors treated with AE6 showed a decrease in E7 expression. In addition, an antisense ODN targeted at the retinoblastoma gene was able to reverse some of the inhibitory effect of AE6 on CaSki cells, indicating that AE6 inhibited E7 synthesis. This study further demonstrates that anti-HPV ODNs may be useful therapeutically.


Subject(s)
Gene Expression Regulation, Viral , Genes, Viral , Papillomaviridae/genetics , Repressor Proteins , Uterine Cervical Neoplasms/virology , Viral Structural Proteins/genetics , Animals , Antineoplastic Agents/pharmacology , Base Sequence , Cell Adhesion , Female , Growth Inhibitors/pharmacology , Humans , In Vitro Techniques , Mice , Mice, Nude , Molecular Sequence Data , Neoplasm Transplantation , Oligonucleotides, Antisense/chemistry , Oligonucleotides, Antisense/pharmacology , Oncogene Proteins, Viral/metabolism , Papillomavirus E7 Proteins , RNA, Messenger/genetics , Retinoblastoma Protein/metabolism , Tumor Cells, Cultured , Uterine Cervical Neoplasms/pathology
8.
J Gen Virol ; 75 ( Pt 10): 2663-70, 1994 Oct.
Article in English | MEDLINE | ID: mdl-7931152

ABSTRACT

The transforming genes E6 and E7 of human papillomavirus (HPV) type 16 and other HPV types are expressed from a bicistronic mRNA with a characteristic spacing of 3 to 6 bp between the termination codon of E6 and the initiation codon of E7. Plasmid pSP64E6E7 which contains the reading frames of both E6 and E7 was constructed in order to study the expression of both proteins in a coupled transcription/rabbit reticulocyte translation system. Both E6 and E7 proteins were expressed simultaneously. This translation could be interfered with by antisense oligonucleotides corresponding to various regions of the transcript. Antisense oligonucleotides targeted at sequences flanking either side of the translation initiation codon of the E6 open reading frame were effective in inhibiting the synthesis of both proteins, whereas oligonucleotides complementary to the coding regions downstream of the first start codon showed either a considerably reduced effect or none at all. In particular, there was limited inhibition of E7 translation by antisense oligonucleotides flanking the translation start region of the E7 gene. In the presence of RNase H, it was possible to selectively inhibit the synthesis of either E6 or E7 by several gene-internal antisense oligonucleotides. We conclude that HPV16 E6-E7 bicistronic mRNA is fully functional and that both proteins are translated with equal efficiency via the scanning mechanisms with reinitiation at the second open reading frame. In addition, both AE6 and AE7 may have therapeutical potential as they are capable of inhibiting the proliferation of CaSki cells which contain the HPV16 genome.


Subject(s)
Genes, Viral , Oncogene Proteins, Viral/biosynthesis , Papillomaviridae/genetics , Protein Biosynthesis , RNA, Messenger/metabolism , Repressor Proteins , Viral Structural Proteins/genetics , Amino Acid Sequence , Animals , Base Sequence , Cell Line , DNA Primers , Female , Humans , Molecular Sequence Data , Oligodeoxyribonucleotides , Oligonucleotides, Antisense/pharmacology , Papillomaviridae/drug effects , Papillomaviridae/physiology , Papillomavirus E7 Proteins , Plasmids , Polymerase Chain Reaction , RNA, Viral/metabolism , Rabbits , Reticulocytes/metabolism , Transcription, Genetic , Tumor Cells, Cultured , Uterine Cervical Neoplasms , Virus Replication
9.
AIDS Res Hum Retroviruses ; 8(5): 605-12, 1992 May.
Article in English | MEDLINE | ID: mdl-1515213

ABSTRACT

Short-term (1 h) treatment with a newly synthesized sulfated polysaccharide, curdlan sulfate (CRDS), showed relatively weak blocking effects on the binding of human immunodeficiency virus type 1 (HIV-1) to the surface of H9 cells. To investigate whether long-term treatment with CRDS could strengthen this effect, CRDS in various doses (0.1, 1, 10, and 100 micrograms/ml) was used in 2-week treatment periods in four separate protocols or "Procedures." SF titers and p24 antigen levels were partially suppressed during long-term CRDS treatment but returned to control levels after the treatment was terminated. In addition, no direct cytotoxicity of CRDS to H9 cells or H9/HIV-1 cells was observed in vitro in the course of continuous exposure to 100 micrograms/ml CRDS for 2 weeks. These results demonstrate the effectiveness of long-term treatment of cells infected with HIV-1 in inhibiting virus expression. The most dramatic inhibition results were obtained when the compound was present both at the time of exposure of cells to virus and during a long-term follow-up treatment. These results show that CRDS inhibits both the cell-free and cell-associated transmission of HIV-1 to host cells and interferes with early events in virus infection. In contrast, CRDS exhibits no significant virucidal activity and has little effect on already infected cells.


Subject(s)
Antiviral Agents/pharmacology , Glucans/pharmacology , HIV-1/drug effects , beta-Glucans , Antiviral Agents/toxicity , Base Sequence , Blotting, Southern , Cell Division/drug effects , Cell Line , DNA, Viral , Glucans/toxicity , Humans , Molecular Sequence Data , Polymerase Chain Reaction , Time Factors
10.
AIDS Res Hum Retroviruses ; 7(4): 409-15, 1991 Apr.
Article in English | MEDLINE | ID: mdl-1676899

ABSTRACT

Action mechanisms of a newly synthesized polysaccharide, curdlan sulfate (CRDS), on human immunodeficiency virus type 1 (HIV-1) infection were investigated in vitro using syncytium formation microassay and p24 antigen capture enzyme-linked immunosorbent assay. These assays measured the titer of infectious virions and the amounts of HIV-1 core antigen p24 in soluble, intraviral, and intracellular forms. CRDS treatments were performed for 1 h at 37 degrees C. H9 cells pretreated with 0.1 to 100.0 micrograms/ml of CRDS appreciably inhibited HIV-1 infection. CRDS-treated HIV-1 virions were less able to infect H9 cells than untreated virions. The simultaneous treatment of H9 cells and HIV-1 virions with CRDS induced a significant inhibition of HIV-1 infection, resulting in the temporary disappearance of virions at the highest dose of CRDS. In contrast, CRDS treatment of newly HIV-1-infected H9 cells caused a significant decrease in the titer of infectious HIV-1 and the p24 amounts of all three forms, but no absolute elimination. Taken together, these results indicate that CRDS may block the binding of the HIV-1 envelope to the H9 cell surface, with emphasis on the high affinity of CRDS to the HIV-1 envelope.


Subject(s)
Antiviral Agents/pharmacology , CD4-Positive T-Lymphocytes/microbiology , Glucans/pharmacology , HIV Infections/microbiology , HIV-1/growth & development , Virus Replication/drug effects , beta-Glucans , Cell Line , Enzyme-Linked Immunosorbent Assay , Gene Products, gag/analysis , HIV Core Protein p24 , HIV-1/drug effects , Humans , Viral Core Proteins/analysis , Virion/drug effects
11.
J Virol ; 63(9): 3579-85, 1989 Sep.
Article in English | MEDLINE | ID: mdl-2474670

ABSTRACT

A panel of seven monoclonal antibodies against the relatively conserved CD4-binding domain on human immunodeficiency virus type 1 (HIV-1) gp120 was generated by immunizing mice with purified gp120. These monoclonal antibodies reacted specifically with gp120 in an enzyme-linked immunosorbent assay and Western blots (immunoblots). By using synthetic peptides as antigens in the immunosorbent assay, the epitopes of these seven monoclonal antibodies were mapped to amino acid residues 423 to 437 of gp120. Further studies with radioimmunoprecipitation assays showed that they cross-reacted with both gp120 and gp160 of diverse HIV-1 isolates (HTLV-IIIB, HTLV-IIIRF, HTLV-IIIAL, and HTLV-IIIWMJ). They also bound specifically to H9 cells infected with HTLV-IIIB, HTLV-IIIRF, HTLV-IIIAL, HTLV-IIIZ84, and HTLV-IIIZ34 in indirect immunofluorescence studies. In addition, they blocked effectively the binding of HIV-1 to CD4+ C8166 cells. Despite the similarity of these properties, the monoclonal antibodies differed in neutralizing activity against HTLV-IIIB, HTLV-IIIRF, and HTLV-IIIAL, as demonstrated in both syncytium-forming assays and infectivity assays. Our findings suggest that these group-specific monoclonal antibodies to the putative CD4-binding domain on gp120 are potential candidates for development of therapeutic agents against acquired immunodeficiency disease syndrome.


Subject(s)
Antibodies, Monoclonal/immunology , Antigens, Differentiation, T-Lymphocyte/immunology , Receptors, Virus/immunology , Retroviridae Proteins/immunology , Animals , Antibodies, Monoclonal/biosynthesis , Epitopes/analysis , HIV/immunology , HIV Envelope Protein gp120 , Male , Mice , Mice, Inbred BALB C , Neutralization Tests , Receptors, HIV , Receptors, Virus/analysis
12.
J Virol ; 63(3): 1400-3, 1989 Mar.
Article in English | MEDLINE | ID: mdl-2783739

ABSTRACT

A new DNA polymerase and DNase activity were identified from cells infected with human B-lymphotropic herpesvirus (HBLV). DNA polymerase associated with HBLV infection was similar in its sensitivity to inhibition by ppi analogs as other herpesvirus-specific DNA polymerases but was dissimilar in its inhibition by certain nucleoside triphosphates.


Subject(s)
B-Lymphocytes/microbiology , DNA-Directed DNA Polymerase/physiology , Herpesviridae/enzymology , DNA-Directed DNA Polymerase/isolation & purification , Deoxyribonucleases/metabolism , Humans , Hydrogen-Ion Concentration , Kinetics , Nucleic Acid Synthesis Inhibitors , Nucleotides/pharmacology , Potassium Chloride/pharmacology , Substrate Specificity
13.
Eur J Epidemiol ; 4(4): 426-34, 1988 Dec.
Article in English | MEDLINE | ID: mdl-2904886

ABSTRACT

Serum samples collected from four groups of individuals in the Washington, D.C. area were examined for the presence of IgG and IgM classes of antibody reacting against HTLV-I and HIV-1. These four groups were: (1) healthy adults with negative premarital VDRL test for syphilis (n = 113), (2) miscellaneous common disease patients (n = 155), (3) drug abusers (n = 130), and (4) homosexual men (n = 187). The former two groups are considered to be low-risk groups, and the latter two, high-risk groups. The prevalence of IgG antibody on ELISA/Western blot tests for these groups were respectively: (1) 5.3%/1.8%, (2) 5.2%/1.9%, (3) 13.9%/4.6%, and (4) 4.3%/1.6% for HTLV-I, and (1) 2.7%/0.9%, (2) 4.5%/0%, (3) 12.3%/5.4%, and (4) 8.0%/5.9% for HIV-1. Instances of possible concomitant infection as shown by the presence of antibodies against both HTLV-I and HIV-1 were found only in the latter two high-risk groups, i.e. two (1.5%) in group (3), and three (1.6%) in group (4) as confirmed by both Western blot and immunofluorescence tests. Out of 97 sera collected from drug abusers in 1985-86 which had IgG antibody by Western blot test against HIV-1, 23 (23.7%) were HTLV-I antibody positive by ELISA test (Group 5), and 8 of these were confirmed by Western blot test. Among these 8 persons, IgM antibody against HTLV-I was found in 2, while that against HIV-1 was positive in 7 persons.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
HIV Antibodies/analysis , HIV/immunology , HTLV-I Antibodies/analysis , Human T-lymphotropic virus 1/immunology , Immunoglobulin G/analysis , Immunoglobulin M/analysis , Acquired Immunodeficiency Syndrome/complications , Acquired Immunodeficiency Syndrome/epidemiology , Acquired Immunodeficiency Syndrome/immunology , Adult , Aged , Blotting, Western/methods , Deltaretrovirus Infections/complications , Deltaretrovirus Infections/epidemiology , Deltaretrovirus Infections/immunology , District of Columbia , Enzyme-Linked Immunosorbent Assay , Female , Fluorescent Antibody Technique , Homosexuality , Humans , Injections, Intravenous , Male , Middle Aged , Random Allocation , Retrospective Studies , Substance-Related Disorders/immunology
14.
JAMA ; 255(16): 2167-72, 1986 Apr 25.
Article in English | MEDLINE | ID: mdl-3007789

ABSTRACT

Using blood samples collected since 1978, we investigated the epidemiology of human T-cell lymphotropic virus type III (HTLV-III), the etiologic agent of the acquired immunodeficiency syndrome, in a group of 378 homosexually active men who have resided in New York City since the acquired immunodeficiency syndrome epidemic began. The anti-HTLV-III prevalence was 6.6% in sera from 1978 or 1979, and the subsequent annual incidence of seroconversion among susceptible men ranged between 5.5% and 10.6%. The highest incidences were in recent years, even though these men reported a decrease in their sexual activity during this time. These data demonstrate the continuing risk of HTLV-III infections in the homosexual population studied and emphasize the need for more effective prevention of transmission. The year during which antibody was first present was the only factor identified that was associated with altered cell-mediated immunity in antibody-positive men. Men who became antibody positive in 1981 or earlier currently had significantly lower OKT4/OKT8 ratios than did those who seroconverted more recently. Further follow-up will be necessary to establish the reasons for this association.


Subject(s)
Acquired Immunodeficiency Syndrome/epidemiology , Homosexuality , Retroviridae Infections/epidemiology , Antibodies, Viral/analysis , Deltaretrovirus/immunology , Humans , Male , New York City , Prospective Studies , Sexual Behavior , T-Lymphocytes/classification , Time Factors
15.
Cancer Res ; 45(9 Suppl): 4621s-4623s, 1985 Sep.
Article in English | MEDLINE | ID: mdl-2990696

ABSTRACT

Sera from 50 Japanese hemophiliacs were screened for antibodies to human T-lymphotropic retrovirus types I and III (HTLV-I and -III). As a whole, antibody to HTLV-I, antibody to HTLV-III, and antibodies to HTLV-I and -III were detected in sera from 2, 17, and 6 hemophiliacs, respectively. Among them, two hemophiliacs developed acquired immunodeficiency syndrome who were positive for both antibodies to HTLV-I and -III in sera. All of the others were asymptomatic. Most of the blood products transfused into these hemophiliacs were imported from abroad, whence the source of HTLV-III infection presumably originated. However, since quite a high percentage of these antibody-positive hemophiliacs was positive for antibody to HTLV-I, even though they are native residents in HTLV-I nonendemic areas of Japan, some special factors may have participated in HTLV-I infection. These special factors should be investigated in the future.


Subject(s)
Antibodies, Viral/analysis , Hemophilia A/immunology , Acquired Immunodeficiency Syndrome/immunology , Adolescent , Adult , Blood Transfusion , Child , Child, Preschool , Factor IX/therapeutic use , Factor VIII/therapeutic use , Female , HIV Antibodies , Hemophilia A/blood , Humans , Hypersensitivity, Delayed , Immunoglobulins/analysis , Japan , Killer Cells, Natural/immunology , Leukocyte Count , Lymphocyte Activation , Male , Middle Aged , Retroviridae Infections/immunology
19.
Int J Cancer ; 33(2): 161-5, 1984 Feb 15.
Article in English | MEDLINE | ID: mdl-6319298

ABSTRACT

Mouse monoclonal antibody to HTLV p19 was used to locate HTLV p19 on the surface of cells and virions by immunofluorescence microscopy (IFM) and immunoelectron microscopy (IEM). When HTLV-producing cells HUT 102 (B2 clone), MT-2 and strain A were used as target cells, HTLV p19 was detected on the surface of cells and virions as spots or small sectors by both IFM and IEM. Cells infected with animal type-C retroviruses, e.g., gibbon ape leukemia virus, simian sarcoma virus, feline leukemia virus, and Gross murine leukemia virus, were completely negative for HTLVp19 expression. Other human T cells not producing HTLV, including HUT78 and HSB2-0, immature or pre-T cells (Molt-3) derived from leukemia patients, and fresh peripheral blood T cells from healthy persons, were also negative. In addition, B cells including Rob-B, IM-9, Raji, and BT-1 did not react with the monoclonal antibody to HTLV p19. In the light of the presence of HTLV p19 in the periphery of acetone-fixed HTLV-producing cells as shown by IFM, it seems most likely that HTLV p19 is an internal antigen of HTLV with part of its structure protruding out of the viral and cell membrane. The monoclonal antibody to HTLV p19 did not lyse HTLV-producing cells in the presence of complement, as expected, because the antibody is an IgG1. Antibody-dependent cell-mediated cytotoxicity was also studied by the 51Cr-release assay. No cytotoxicity was observed. Although HTLV p19 does not contribute to the destruction of malignant T cells for treatment and/or virions for prophylaxis, this protein is an important marker for diagnosis of HTLV infection. The patterns of HTLV p19 expression described above were exactly the same for American HTLV-producing HUT 102 (B2 clone), for strain A cells and for Japanese HTLV-producing MT-2 cells. These results further substantiate the close relationship of the Japanese and American HTLV isolates.


Subject(s)
Antibodies, Monoclonal/immunology , Antigens, Viral/analysis , Cell Membrane/immunology , Deltaretrovirus/immunology , Retroviridae/immunology , Virion/immunology , Clone Cells , Cytotoxicity, Immunologic , Humans , Microscopy, Electron , Microscopy, Fluorescence , Virus Replication
20.
J Natl Cancer Inst ; 69(2): 435-41, 1982 Aug.
Article in English | MEDLINE | ID: mdl-6287077

ABSTRACT

Cultured human hematopoietic cells from several normal and leukemic sources, including those cells initiated after exposure to primate type C retroviruses were tested for their capacity to induce tumors in young athymic BALB/c (nu/nu) mice after sc inoculation. An attempt was made to correlate these results with virus expression and chromosome patterns. Progressively growing tumor formation was observed in 5 of 18 normal diploid B-lymphoblast lines from normal peripheral blood and in one of three diploid B-lymphoblast lines from leukemic donors established after infection with primate type C viruses (gibbon ape leukemia virus or simian sarcoma virus). In contrast, none of eight spontaneously transformed B-lymphoblast lines with normal diploid karyotypes formed progressively growing tumors, although one formed a tumor that remained the same size (0.5 cm) for several months. Progressive tumor formation occurred in four of seven previously established cell lines of different cell types that had abnormal karyotypes. Of the normal diploid B-lymphoblast cultures exposed to type C viruses, 12 were tested for the presence of viral RNA and structural proteins (p12, p30, gp70), and this information was correlated with tumorigenicity. Four of the six cultures expressing viral RNA or proteins were tumorigenic, whereas only one of six cultures that did not express virus information was positive. The results of this study suggest that expression of type C viral RNA and proteins by human B-lymphoblasts increases their tumorigenicity in nude mice. It is also apparent that caution must be used in attempts to correlate cell tumorigenicity and chromosome abnormalities in nude mice.


Subject(s)
Cell Transformation, Viral , Neoplasms, Experimental/etiology , Retroviridae , Animals , B-Lymphocytes , Cell Line , Cells, Cultured , Chromosome Aberrations , Hematopoietic Stem Cells/ultrastructure , Humans , Karyotyping , Mice , Mice, Nude , Neoplasm Transplantation , Neoplasms, Experimental/ultrastructure
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