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10. Combination Therapy with Zidovudine Prevents Selection of Human Immunodeficiency Virus Type 1 Variants Expressing High-Level Resistance to L-697,661, a Nonnucleoside Reverse Transcriptase Inhibitor

Author

Staszewski, S., Massari, F. E., Kober, A., Göhler, R., Durr, S., Anderson, K. W., Schneider, C. L., Waterbury, J. A., Bakshi, K. K., Taylor, V. I., Hildebrand, C. S., Kreisl, C., Hoffstedt, B., Schleif, W. A., von Briesen, H., Rübsamen-Waigmann, H., Calandra, G. B., Ryan, J. L., Stille, W., Emini, E. A., and Byrnes, V. W.

Published
1995

13. Cryopreservation of human mucosal leukocytes

Author

Hughes, SM, Shu, Z, Levy, CN, Ferre, AL, Hartig, H, Fang, C, Lentz, G, Fialkow, M, Kirby, AC, Waldorf, KMA, Veazey, RS, Germann, A, Von Briesen, H, McElrath, MJ, Dezzutti, CS, Sinclair, E, Baker, CAR, Shacklett, BL, Gao, D, Hladik, F, Hughes, SM, Shu, Z, Levy, CN, Ferre, AL, Hartig, H, Fang, C, Lentz, G, Fialkow, M, Kirby, AC, Waldorf, KMA, Veazey, RS, Germann, A, Von Briesen, H, McElrath, MJ, Dezzutti, CS, Sinclair, E, Baker, CAR, Shacklett, BL, Gao, D, and Hladik, F

Abstract

Background: Understanding how leukocytes in the cervicovaginal and colorectal mucosae respond to pathogens, and how medical interventions affect these responses, is important for developing better tools to prevent HIV and other sexually transmitted infections. An effective cryopreservation protocol for these cells following their isolation will make studying them more feasible. Methods and Findings: To find an optimal cryopreservation protocol for mucosal mononuclear leukocytes, we compared cryopreservation media and procedures using human vaginal leukocytes and confirmed our results with endocervical and colorectal leukocytes. Specifically, we measured the recovery of viable vaginal T cells and macrophages after cryopreservation with different cryopreservation media and handling procedures. We found several cryopreservation media that led to recoveries above 75%. Limiting the number and volume of washes increased the fraction of cells recovered by 10-15%, possibly due to the small cell numbers in mucosal samples. We confirmed that our cryopreservation protocol also works well for both endocervical and colorectal leukocytes. Cryopreserved leukocytes had slightly increased cytokine responses to antigenic stimulation relative to the same cells tested fresh. Additionally, we tested whether it is better to cryopreserve endocervical cells on the cytobrush or in suspension. Conclusions: Leukocytes from cervicovaginal and colorectal tissues can be cryopreserved with good recovery of functional, viable cells using several different cryopreservation media. The number and volume of washes has an experimentally meaningful effect on the percentage of cells recovered. We provide a detailed, step-by-step protocol with best practices for cryopreservation of mucosal leukocytes.

Published
2016

14. Selection of HIV-1 genotypes by cultivation in different primary cells

Author

Unger Re, Ursula Dietrich, Panhans B, Becker K, H. Rübsamen-Waigmann, von Briesen H, I. Raudonat, Manuel Grez, and H. Ruppach

Subjects
Adult, Male, Virus Cultivation, Lymphocyte, Molecular Sequence Data, Immunology, HIV Infections, Viral quasispecies, HIV Envelope Protein gp120, Biology, Lymphocyte Activation, Peripheral blood mononuclear cell, Monocytes, Virus, law.invention, law, medicine, Humans, Immunology and Allergy, Amino Acid Sequence, Lymphocytes, Cells, Cultured, Polymerase chain reaction, Macrophages, Monocyte, Genetic Variation, Virology, Infectious Diseases, medicine.anatomical_structure, Cell culture, HIV-1, Leukocytes, Mononuclear, Female
Abstract

Objectives: To determine the representation of particular HIV-1 genotypes during cultivation in different primary cell-culture systems compared with the spectrum of the quasispecies in vivo. Methods: Primary isolates of HIV-1 were recovered by isolation in cultures of lymphocytes, mixed mononuclear cells (MNC), and monocytes/macrophages. Nucleotide sequence determination of the C2-V3 region of gp120 of HIV was performed on 10-20 independently isolated clones derived by polymerase chain reaction from the culture systems, the uncultured peripheral blood MNC (PBMC) as well as plasma. Results: Several predominant HIV genotypes were found in the uncultured PBMC from each of the patients. The most frequent genotypes in PBMC were also the most frequent types in plasma. In addition, lymphocytes, macrophages or mixed MNC cultures allowed the outgrowth of variants that were underrepresented in uncultured PBMC. We showed that the virus cultivation systems used in this study selected differently for the genetic variants. Whereas some genotypes were present in all three culture systems, although at different frequencies, others were exclusively found in a specific culture system. Conclusions: These results demonstrate that monocyte/macrophage and mixed MNC culture systems complement the standard lymphocyte culture in terms of the spectrum of genotypically different virus variants obtained in vitro.

Published
1999

16. Standard Conditions of Virus Isolation Reveal Biological Variability of HIV Type 1 in Different Regions of the World

Author

Björndal A, Etienne Karita, Tomlinson P, Shahin Ranjbar, Esser R, von Briesen H, Harvey Holmes, Bette T. Korber, Bernardo Galvão-Castro, and H. Rübsamen-Waigmann

Subjects
Syncytium, viruses, Immunology, Biology, Virology, Virus, Infectious Diseases, Immune system, Viral replication, Immunity, Genotype, Genetic variation, Seroconversion
Abstract

HIV-1 isolates were obtained from four countries within the framework of the WHO Network for HIV Isolation and Characterization. The use of standard HIV isolation procedures allowed us to compare the biological properties of 126 HIV-1 isolates spanning five genetic subtypes. In primary isolation cultures, viruses from Uganda and Brazil appeared early and replicated without delay, whereas the replication of Thai viruses was delayed by several weeks. Regardless of genetic subtype or country of origin, blood samples collected more than 2 years after seroconversion yielded virus that replicated efficiently in the primary isolation cultures. None of the isolates obtained from Thailand or Rwanda replicated in cell lines, whereas 5 of the 13 Brazilian isolates and 7 of the 11 Ugandan isolates replicated and induced syncytia in MT-2 cells. As expected for virus isolates obtained early in HIV-1 infection (within 2 years of seroconversion), all viruses from Brazil, Rwanda, and Thailand showed a slow/low replicative pattern. For the Ugandan samples, the time from seroconversion was known precisely for a few of the samples and only in one case was less than 2 years. This may explain why the five viruses that were able to replicate in all cell lines, and thus classified as rapid/high, were of Ugandan origin. Viruses able to induce syncytia in MT-2 cells, also induced syncytia in PBMC. However, 8 slow/low viruses (out of 27) gave discordant results, inducing syncytia in PBMC but not in MT-2 cells. Furthermore, using syncytium induction as a marker, changes in virus populations during early in vitro passage in PBMC could be observed

Published
1994

21. HIV-1 and HIV-2 infections in a high-risk population in Bombay, India: evidence for the spread of HIV-2 and presence of a divergent HIV-1 subtype

Author

Pfützner A, Ursula Dietrich, von Eichel U, von Briesen H, Hd, Brede, Jk, Maniar, and Rübsamen-Waigmann H

Subjects
Male, Acquired Immunodeficiency Syndrome, Sequence Homology, Amino Acid, Urban Population, HIV Seropositivity, HIV-2, Molecular Sequence Data, HIV-1, Humans, India, Female, Amino Acid Sequence, HIV Envelope Protein gp120
Abstract

A high-risk population (patients of a sexually transmitted disease clinic and the GT hospital in Bombay) was tested for antibodies against HIV-1 and HIV-2. Among 405 serum samples, 226 had previously been classified HIV-positive in India using different locally available enzyme-linked immunosorbent assay (ELISA) tests. The serology of 179 samples was unknown. All 405 samples were tested at the Georg-Speyer-Haus (GSH) with the Pasteur HIV-1/2-Combi-ELISA. Positive samples were further analyzed with HIV-1 and HIV-2 Western blot kits from Dupont and Pasteur, respectively. A very high seroprevalence of HIV was found in this population. Among the 179 unscreened samples, 69 (38.5%) were positive in the ELISAs as well as the Western blots for HIV-1 or HIV-2. Among the prescreened samples, only 174 (77%) were confirmed HIV-positive. Altogether, 243 of 405 sera were HIV-positive. Of these, 184 (76%) were reactive with HIV-1, 10 (4%) were reactive with HIV-2, and 49 (20%) had dual reactivity to HIV-1 and HIV-2. Previous data from the Indian Council of Medical Research had already suggested a possible high prevalence of HIV-1 in India. Our results confirm this view. The finding of a substantial spread of HIV-2 infection was, however, totally unexpected in India, but confirms our previous study which had already demonstrated the existence of HIV-2 in this country. Asia can thus no longer be considered free of HIV-2, and testing for HIV-2 appears mandatory, at least in India.(ABSTRACT TRUNCATED AT 250 WORDS)

Published
1992

24. Implications of HIV variability for transmission scientific and policy issues. Expert group of the Joint United Nations Programme on HIV/AIDS

Author

Anderson, R, Barre-Sinoussi, F, Bradac, J, Burke, DS, Essex, M, Fenyo, EM, Galvao-Castro, B, Hu, DJ, Jaffe, H, Karamov, E, Kuiken, C, Kunanusont, C, Laga, M, Lower, J, Mastro, T, McCutchan, FE, Opio, A, Pauli, G, Sutherland, D, Schwartlander, B, de Vincenzi, I, von Briesen, H, Wasi, C, Weber, J, Piot, P, Carael, M, Osmanov, S, Anderson, R, Barre-Sinoussi, F, Bradac, J, Burke, DS, Essex, M, Fenyo, EM, Galvao-Castro, B, Hu, DJ, Jaffe, H, Karamov, E, Kuiken, C, Kunanusont, C, Laga, M, Lower, J, Mastro, T, McCutchan, FE, Opio, A, Pauli, G, Sutherland, D, Schwartlander, B, de Vincenzi, I, von Briesen, H, Wasi, C, Weber, J, Piot, P, Carael, M, and Osmanov, S

Published
1997

27. Molecular cloning and analysis of functional envelope genes from human immunodeficiency virus type 1 sequence subtypes A through G. The WHO and NIAID Networks for HIV Isolation and Characterization

Author

Gao, F, primary, Morrison, S G, additional, Robertson, D L, additional, Thornton, C L, additional, Craig, S, additional, Karlsson, G, additional, Sodroski, J, additional, Morgado, M, additional, Galvao-Castro, B, additional, von Briesen, H, additional, Beddows, S, additional, Weber, J, additional, Sharp, P M, additional, Shaw, G M, additional, and Hahn, B H, additional

Published
1996
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42. Establishment of two Epstein-Barr virus negative Burkitt cell lines from a patient with AIDS and B-cell lymphoma

Author

Ganser, A, Carlo-Stella, C, Bartram, CR, Boehm, T, Heil, G, Henglein, B, Muller, H, Raghavachar, A, von Briesen, H, and Griesinger, F

Abstract

To analyze the pathogenesis of B-cell lymphomas in patients with acquired immunodeficiency syndrome (AIDS), we studied two cell lines, Es I and Es III, established from one such lymphoma for the presence of sequences of the Epstein-Barr virus (EBV) and the human immunodeficiency virus [HIV; lymphadenopathy-associated virus (LAV/HTLV- III)] as well as for the presence of cytogenetic abnormalities and monoclonal rearrangements of immunoglobulin and T-cell receptor genes. Both cell lines expressed the same IgM, kappa phenotype as the original lymphoma. The karyotype of Es I was 46, XY, t(8;14), 2 p+, inv (6p), 17p-, and the cells of Es III had an additional i(7q). Immunoglobulin gene studies demonstrated the identical monoclonal rearrangements in both cell lines. Neither EBV nor HIV sequences were detectable in the malignant B cells at the genomic level, leading to the conclusion that mechanisms other than transformation by EBV or HIV may have contributed to the B-cell lymphoma in this patient and possibly also to the generally increased frequency in patients with AIDS.

Published
1988
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43. Synthesis and in vitroAntiviral Properties of Amphiphilic Dinucleoside Phosphate Derivatives of 2′,3-dideoxycytidine (ddC)

Author

Schott, H., Häussler, M. P., Gowland, P., Bender, A., von Briesen, H., and Schwendener, R. A.

Abstract

N4-hexadecyl-5′-0-(4-monomethoxytrityl)-2′-deoxycytidine-3′-hydrogenphosphonate and 5′-0-(4-monomethoxytrityl)-2′-deoxythymidine-3′-0-hydrogenphosphonate were condensed with 2′,3′-dideoxycytidine (ddC) according to the hydrogenphosphonate method to yield N4-hexadecyl-2′-deoxycytidylyl-(3′-5′)-2′,3′-dideoxycytidine (N4-hexadecyldC-ddC) and 2′-deoxythymidylyl-(3′-5′)-N4-palmitoyl-2′,3′-dideoxycytidine (dT-N4-palmddC). N4-palmitoyl-2′,3′-dideoxycytidine (N4-palmddC) was synthesized by reacting palmitic anhydride with ddC. Both dinucleoside phosphates have amphiphilic properties and represent a new class of ddC derivatives in which in the case of the dinucleosides, the ddC-5′-monophosphate is masked with lipophilic residues of variable stability. The ddC derivatives can be solubilized in water by micelle formation and, because they have lipophilic residues, they can be incorporated into the lipid membranes of liposomes. The ddC derivatives were shown to have antiviral activities comparable to those of AZT and ddC when tested in vitroagainst HIV-1-infected HeLa and H9 cells as well as infected human monocytes/macrophages.

Published
1995
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44. Molecular cloning of two west African human immunodeficiency virus type 2 isolates that replicate well in macrophages: a Gambian isolate, from a patient with neurologic acquired immunodeficiency syndrome, and a highly divergent Ghanian isolate.

Author

Kühnel, H, von Briesen, H, Dietrich, U, Adamski, M, Mix, D, Biesert, L, Kreutz, R, Immelmann, A, Henco, K, and Meichsner, C

Abstract

Human immunodeficiency virus type 2 (HIV-2)-related viruses were isolated from a Gambian dying of exclusively neurological disease (HIV-2D194) and from an asymptomatic Ghanian (HIV-2D205). Both strains exhibited properties of HIV-1 biological subtype c: they grew slowly and induced few or no syncytia but eventually produced high levels of particle-associated reverse transcriptase in cultures of fresh peripheral blood lymphocytes, and they established stable infection of T-lymphoma (HUT-78) and monocytic (U937) cell lines. Each produced even higher levels of reverse transcriptase when fresh human monocytes/macrophages were used as target cells. The viruses were molecularly cloned after a single passage in culture, in order to minimize in vitro selection of subtypes present in vivo. Restriction-site analysis showed heterogeneity within each isolate. Nucleotide sequence analysis of a portion of the HIV-2D194 genome revealed that it is a member of the prototypic HIV-2 family, displaying 13% divergence versus HIV-2ROD and HIV-2NIHZ, as compared to 9% divergence between HIV-2ROD and HIV-2NIHZ. In contrast, HIV-2D205 is the most highly divergent HIV-2 strain yet described: it is equidistant in relation between the known HIV-2 strains and the simian immunodeficiency virus isolates from rhesus macaque monkeys (23-25% divergence).

Published
1989
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45. Measurement of Electroproduction of Muon Pairs

Author

Earles, D. R., primary, Chase, R C., additional, Faissler, W. L., additional, Gettner, M. W., additional, Glass, G., additional, Lutz, G., additional, Parsons, R. G., additional, Rothwell, P. L., additional, Moy, K. M., additional, von Briesen, H., additional, von Goeler, E., additional, and Weinstein, Roy, additional

Published
1970
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50. Secretion potential of HIV-infected human monocytes/macrophages

Author

Esser, R., von Briesen, H., Brugger, W., Ceska, M., Liehl, E., Rubsamen-Waigmann, H., and Andreesen, R.

Subjects
HIV infection -- Research, Monocytes -- Research, Macrophages -- Research
Abstract

AUTHORS: R. Esser(1), H. von Briesen(1), W. Brugger(2), M. Ceska(3), E. Liehl(3), H. Rubsamen-Waigmann(1), and R. Andreesen(2). (1)Georg Speyer Haus, Frankfurt; (2)Medizinische Klinik, Freiburg; and Sandoz Forschunginstitute, Wien, Germany. According [...]

Published
1991

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