Retrovirus particles with type C morphology were found in two T-cell lymphoblastoid cell lines, HUT 102 and CTCL-3, and in fresh peripheral blood lymphocytes obtained from a patient with a cutaneous T-cell lymphoma (mycosis fungoides). The cell lines continuously produce these viruses, which are collectively referred to as HTLV, strain CR(HTLVCR). Originally, the production of virus from HUT 102 cells required induction with 5-iodo-2′-deoxyuridine, but the cell line became a constitutive producer of virus at its 56th passage. Cell line CTCL-3 has been a constitutive producer of virus from its second passage in culture. Both mature and immature extracellular virus particles were seen in thin-section electron micrographs of fixed, pelleted cellular material; on occasion, typical type C budding virus particles were seen. No form of intracellular virus particle has been seen. Mature particles were 100-110 nm in diameter, consisted of an electron-dense core surrounded by an outer membrane separated by an electron-lucent region, banded at a density of 1.16 g/ml on a continuous 25-65% sucrose gradient, and contained 70S RNA and a DNA polymerase activity typical of viral reverse transcriptase (RT; RNA-dependent DNA nucleotidyltransferase). Under certain conditions of assay, HTLVCR RT showed cation preference for Mg2+ over Mn2+, distinct from the characteristics of cellular DNA polymerases purified from human lymphocytes and the RT from most type C viruses. Antibodies to cellular DNA polymerase γ and anti-bodies against RT purified from several animal retroviruses failed to detectably interact with HTLVCR RT under conditions that were positive for the respective homologous DNA polymerase, demonstrating a lack of close relationship of HTLVCR RT to cellular DNA polymerases γ or RT of these viruses. Six major proteins, with sizes of approximately 10,000, 13,000, 19,000, 24,000, 42,000, and 52,000 daltons, were apparent when doubly banded, disrupted HTLVCR particles were chromatographed on a NaDodSO4/polyacrylamide gel. The number of these particle-associated proteins is consistent with the expected proteins of a retrovirus, but the sizes of some are distinct from those of most known retroviruses of the primate subgroups.

https://www.pnas.org/content/pnas/77/12/7415.full.pdf

Detection and isolation of type C retrovirus particles from fresh and cultured lymphocytes of a patient with cutaneous T-cell lymphoma

Acquired immune deficiency syndrome (AIDS) is characterized by opportunistic infections and by 'opportunistic neoplasms' (for example, Kaposi's sarcoma). Persistent generalized lymphadenopathy (PGL) is epidemiologically associated with AIDS, especially in male homosexuals. A subset of T lymphocytes positive for the CD4 antigen (also termed T4 antigen), is depleted in AIDS and PGL patients. A retrovirus found in T-cell cultures from these patients is strongly implicated in the aetiology of AIDS because of the high frequency of isolation and the prevalence of specific antibodies in the patients. Here we have detected cell-surface receptors for the AIDS retrovirus (human T-cell leukaemia virus-III (HTLV-III) and lymphadenopathy-associated virus-1 (LAV-1) isolates) by testing the susceptibility of cells to infection with pseudotypes of vesicular stomatitis virus bearing retroviral envelope antigens, and by the formation of multinucleated syncytia on mixing virus-producing cells with receptor-bearing cells. Receptors were present only on cells expressing CD4 antigen; among 155 monoclonal antibodies tested, each of the 14 anti-CD4 antibodies inhibited formation of syncytia and blocked pseudotypes. Productive infection of CD4+ cells with HTLV-III or LAV-1 markedly reduced cell-surface expression of CD4. In contrast, receptors for HTLV-I and HTLV-II were not restricted to CD4+ cells, were not blocked by anti-CD4 antibodies; cells productively infected with HTLV-I and HTLV-II expressed surface CD4. Hence, we conclude that the CD4 antigen is an essential and specific component of the receptor for the causative agent of AIDS.

The CD4 (T4) antigen is an essential component of the receptor for the AIDS retrovirus

Peripheral blood lymphocytes from patients with the acquired immunodeficiency syndrome (AIDS) or with signs or symptoms that frequently precede AIDS (pre-AIDS) were grown in vitro with added T-cell growth factor and assayed for the expression and release of human T-lymphotropic retroviruses (HTLV). Retroviruses belonging to the HTLV family and collectively designated HTLV-III were isolated from a total of 48 subjects including 18 of 21 patients wih pre-AIDS, three of four clinically normal mothers of juveniles with AIDS, 26 of 72 adult and juvenile patients with AIDS, and from one of 22 normal male homosexual subjects. No HTLV-III was detected in or isolated from 115 normal heterosexual subjects. The number of HTLV-III isolates reported here underestimates the true prevalence of the virus since many specimens were received in unsatisfactory condition. Other data show that serum samples from a high proportion of AIDS patients contain antibodies to HTLV-III. That these new isolates are members of the HTLV family but differ from the previous isolates known as HTLV-I and HTLV-II is indicated by their morphological, biological, and immunological characteristics. These results and those reported elsewhere in this issue suggest that HTLV-III may be the primary cause of AIDS.

https://science.sciencemag.org/content/sci/224/4648/500.full.pdf

Frequent detection and isolation of cytopathic retroviruses (HTLV-III) from patients with AIDS and at risk for AIDS

A cell system was developed for the reproducible detection of human T-lymphotropic retroviruses (HTLV family) from patients with the acquired immunodeficiency syndrome (AIDS) or with signs or symptoms that frequently precede AIDS (pre-AIDS). The cells are specific clones from a permissive human neoplastic T-cell line. Some of the clones permanently grow and continuously produce large amounts of virus after infection with cytopathic (HTLV-III) variants of these viruses. One cytopathic effect of HTLV-III in this system is the arrangement of multiple nuclei in a characteristic ring formation in giant cells of the infected T-cell population. These structures can be used as an indicator to detect HTLV-III in clinical specimens. This system opens the way to the routine detection of HTLV-III and related cytopathic variants of HTLV in patients with AIDS or pre-AIDS and in healthy carriers, and it provides large amounts of virus for detailed molecular and immunological analyses.

https://science.sciencemag.org/content/sci/224/4648/497.full.pdf

Detection, Isolation, and Continuous Production of Cytopathic Retroviruses (HTLV-III) from Patients with AIDS and Pre-AIDS

1. Developmental Defects of the Oral and Maxillofacial Region 2. Abnormalities of Teeth 3. Pulpal and Periapical Disease 4. Periodontal Diseases 5. Bacterial Infections 6. Fungal and Protozoal Diseases 7. Viral Infections 8. Physical and Chemical Injuries 9. Allergies and Immunologic Diseases 10. Epithelial Pathology 11. Salivary Gland Pathology 12. Soft Tissue Tumors 13. Hematologic Disorders 14. Bone Pathology 15. Odontogenic Cysts and Tumors 16. Dermatologic Diseases 17. Oral Manifestations of Systemic Diseases 18. Facial Pain and Neuromuscular Diseases 19. Forensic Dentistry Appendix: Differential Diagnosis of Oral and Maxillofacial Diseases Part 1: Mucosal and Soft Tissue Pathology: Color Changes Part 2: Mucosal and Soft Tissue Pathology: Surface Alterations Part 3: Mucosal and Soft Tissue Pathology: Masses or Enlargements Part 4: Radiographic Pathology Part 5: Pathology of Teeth

Oral and Maxillofacial Pathology

Although HIV-1 is the necessary and sufficient causative agent of AIDS, genetic and environmental factors markedly influence the pace of disease progression. Clinical and experimental evidence suggests that human herpesvirus 6 (HHV-6), a cytopathic T-lymphotropic agent, may act as an accelerating factor in the progression of HIV disease, although conclusive in vivo evidence has yet to be attained. To evaluate the effect of HHV-6A on the course of AIDS in a relevant model system, we infected pig-tailed macaques (M. nemestrina) either with HHV-6A (strain GS) or with a pathogenic SIV strain (smE660), or with both viruses. Extensive longitudinal virologic, immunologic and clinical follow-up demonstrated that HHV-6A coinfection dramatically accelerated the progression toward full-blown AIDS. Rapid disease development in coinfected animals was associated with an early depletion of both CD4+ and CD8+ T cells. Simultaneous replication of both viruses was documented in coinfected lymph node tissue. These data establish a new animal model for the study of HHV-6 infection and provide the first conclusive in vivo evidence that HHV-6A acts as a cofactor in the progression of primate immunodeficiency virus disease.

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Accelerated progression to AIDS in macaques coinfected with simian immunodeficiency virus and human herpesvirus 6A

SummaryFeline embryo fibroblast cells were found to be very sensitive to transformation by the primate RNA tumor viruses, simian sarcoma virus (SiSV-1) and HL23V, a virus isolated from the leukemic myelo-blasts of an AML patient. In vitro transformation of feline embryo fibroblast cell cultures with these viruses was 2 to 100 x more efficient than when performed on the normal rat kidney (NRK) or rat embryo fibroblast cell lines commonly used for SiSV-1 transformation studies. Several cell lines of marmoset and human origin were also observed to be less sensitive to transformation by these viruses. Furthermore focus development on feline embryonic fibroblast cultures required only 3-4 days compared to 7-10 days needed using rat or primate cultures.

An in Vitro Transformation Assay for SiSV-1 and HL23V Using Feline Embryonic Fibroblasts

Species-specific transformation of T cells by HVMNE

The present invention relates to the HIV-1 strains MN-ST1 and BA-L which are typical United States HIV-1 isotypes. The present invention relates to DNA segments encoding the envelope protein of MN-ST1 or BA-L, to DNA constructs containing such DNA segments and to host cells transformed with such contructs. The viral isolates and envelope proteins of the present invention are of value for use in vaccines and bioassays for the detection of HIV-1 infection in biological samples, such as blood bank samples.

Molecular clones of hiv-1 and uses thereof

Cette invention concerne les souches MN-ST1 et BA-L de VIH-1 qui sont des isotypes de VIH-1 specifiques a la recherche aux Etats-Unis. L'invention concerne des segments d'ADN codant la proteine d'enveloppe de MN-ST1 ou BA-L, des structures d'ADN contenant de tels segments et des cellules hotes transformees par des structures de ce type. Les isolats viraux et les proteines d'enveloppe de cette invention sont valablement utilisables dans des vaccins et des dosages biologiques permettant de detecter la contamination par le VIH-1 dans des echantillons biologiques, tels que les echantillons d'une banque de sang.

Phillip D. Markham

Papers

Accelerated progression to AIDS in macaques coinfected with simian immunodeficiency virus and human herpesvirus 6A

An in Vitro Transformation Assay for SiSV-1 and HL23V Using Feline Embryonic Fibroblasts

Species-specific transformation of T cells by HVMNE

Molecular clones of hiv-1 and uses thereof

Clones moleculaires de vih-1 et utilisation de ces derniers