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Dive into the research topics where Wolfgang Hofmann is active.

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Featured researches published by Wolfgang Hofmann.


Journal of Biological Chemistry | 1999

Enhanced expression, native purification, and characterization of CCR5, a principal HIV-1 coreceptor.

Tajib Mirzabekov; Norbert Bannert; Michael Farzan; Wolfgang Hofmann; Peter Kolchinsky; Lijun Wu; Richard T. Wyatt; Joseph Sodroski

Seven-transmembrane segment, G protein-coupled receptors (GPCRs) play important roles in many biological processes in which pharmaceutical intervention may be useful. High level expression and native purification of GPCRs are important steps in the biochemical and structural characterization of these molecules. Here, we describe enhanced mammalian cell expression and purification of a codon-optimized variant of the chemokine receptor CCR5, a GPCR that plays a central role in the entry of the human immunodeficiency virus-1 (HIV-1) into immune cells. CCR5 could be solubilized in its native state as determined by its ability to be precipitated by 2D7, a conformation-dependent anti-CCR5 antibody, and by the HIV-1 gp120 envelope glycoprotein. The 2D7 antibody recognized immature and mature forms of CCR5 equally, whereas gp120 preferentially recognized the mature form, a result that underscores a role for posttranslational modification of CCR5 in its HIV-1 coreceptor function. The methods described herein contribute to the analysis of CCR5 and are likely to be applicable to many other GPCRs.


Journal of Virology | 2000

Importance of Membrane Fusion Mediated by Human Immunodeficiency Virus Envelope Glycoproteins for Lysis of Primary CD4-Positive T Cells

Jason A. LaBonte; Trushar Patel; Wolfgang Hofmann; Joseph Sodroski

ABSTRACT In established T-cell lines, the membrane-fusing capacity of the human immunodeficiency virus type 1 (HIV-1) envelope glycoproteins mediates cytopathic effects, both syncytium formation and single-cell lysis. Furthermore, changes in the HIV-1 envelope glycoproteins are responsible for the increased CD4+ T-cell-depleting ability observed in infected monkeys upon in vivo passage of simian-human immunodeficiency virus (SHIV) chimeras. In this study, a panel of SHIV envelope glycoproteins and their mutant counterparts defective in membrane-fusing capacity were expressed in primary human CD4+ T cells. Compared with controls, all of the functional HIV-1 envelope glycoproteins induced cell death in primary CD4+ T-cell cultures, whereas the membrane fusion-defective mutants did not. Death occurred almost exclusively in envelope glycoprotein-expressing cells and not in bystander cells. Under standard culture conditions, most dying cells underwent lysis as single cells. When the cells were cultured at high density to promote syncytium formation, the envelope glycoproteins of the passaged, pathogenic SHIVs induced more syncytia than those of the respective parental SHIV. These results demonstrate that the HIV-1 envelope glycoproteins induce the death of primary CD4+ T lymphocytes by membrane fusion-dependent processes.


Journal of Virology | 2004

Simian Virus 40-Based Replication of Catalytically Inactive Human Immunodeficiency Virus Type 1 Integrase Mutants in Nonpermissive T Cells and Monocyte-Derived Macrophages

Richard Lu; Noriko Nakajima; Wolfgang Hofmann; Monsef Benkirane; Kuan Teh-Jeang; Joseph Sodroski; Alan Engelman

ABSTRACT Integrase function is required for retroviral replication in most instances. Although certain permissive T-cell lines support human immunodeficiency virus type 1 (HIV-1) replication in the absence of functional integrase, most cell lines and primary human cells are nonpermissive for integrase mutant growth. Since unintegrated retroviral DNA is lost from cells following cell division, we investigated whether incorporating a functional origin of DNA replication into integrase mutant HIV-1 might overcome the block to efficient gene expression and replication in nonpermissive T-cell lines and primary cells. Whereas the Epstein-Barr virus (EBV) origin (oriP) did little to augment expression from an integrase mutant reporter virus in EBV nuclear antigen 1-expressing cells, simian virus 40 (SV40) oriT dramatically enhanced integrase mutant infectivity in T-antigen (Tag)-expressing cells. Incorporating oriT into the nef position of a full-length, integrase-defective virus strain yielded efficient replication in Tag-expressing nonpermissive Jurkat T cells without reversion to an integration-competent genotype. Adding Tag to integrase mutant-oriT viruses yielded 11.3-kb SV40-HIV chimeras that replicated in Jurkat cells and primary monocyte-derived macrophages. Real-time quantitative PCR analyses of Jurkat cell infections revealed that amplified copies of unintegrated DNA likely contributed to SV40-HIV integrase mutant replication. SV40-based HIV-1 integrase mutant replication in otherwise nonpermissive cells suggests alternative approaches to standard integrase-mediated retroviral gene transfer strategies.


Nature | 1997

CCR3 and CCR5 are co-receptors for HIV-1 infection of microglia.

Jianglin He; Youzhi Chen; Michael Farzan; Hyeryun Choe; Asa Ohagen; Suzanne Gartner; Jorge Busciglio; Xiaoyu Yang; Wolfgang Hofmann; Walter Newman; Charles R. Mackay; Joseph Sodroski; Dana Gabuzda


Journal of Experimental Medicine | 1997

Two Orphan Seven-Transmembrane Segment Receptors Which Are Expressed in CD4-positive Cells Support Simian Immunodeficiency Virus Infection

Michael Farzan; Hyeryun Choe; Kathleen A. Martin; Luisa Marcon; Wolfgang Hofmann; Gunilla B. Karlsson; Ying Sun; Peter Barrett; Nathalie Marchand; Nancy J. Sullivan; Norma P. Gerard; Craig Gerard; Joseph Sodroski


Journal of Virology | 1995

Replicative function and neutralization sensitivity of envelope glycoproteins from primary and T-cell line-passaged human immunodeficiency virus type 1 isolates.

N Sullivan; Ying Sun; John Li; Wolfgang Hofmann; Joseph Sodroski


Journal of Virology | 1999

Species-Specific, Postentry Barriers to Primate Immunodeficiency Virus Infection

Wolfgang Hofmann; David Schubert; Jason A. LaBonte; Linda Munson; Susan V. Gibson; Jonathan G. Scammell; Paul Ferrigno; Joseph Sodroski


Journal of Virology | 1998

Tat Protein Induces Human Immunodeficiency Virus Type 1 (HIV-1) Coreceptors and Promotes Infection with both Macrophage-Tropic and T-Lymphotropic HIV-1 Strains

Lili Huang; Irene Bosch; Wolfgang Hofmann; Joseph Sodroski; Arthur B. Pardee


Journal of Virology | 1999

Intrinsic Human Immunodeficiency Virus Type 1 Resistance of Hematopoietic Stem Cells Despite Coreceptor Expression

Hongmei Shen; Tao Cheng; Frederick Preffer; David Dombkowski; Michael H. Tomasson; David E. Golan; Otto O. Yang; Wolfgang Hofmann; Joseph Sodroski; Andrew D. Luster; David T. Scadden


Journal of Virology | 1999

Changes in Human Immunodeficiency Virus Type 1 Envelope Glycoproteins Responsible for the Pathogenicity of a Multiply Passaged Simian-Human Immunodeficiency Virus (SHIV-HXBc2)

Mark J. Cayabyab; Gunilla B. Karlsson; Bijan Etemad-Moghadam; Wolfgang Hofmann; Tavis Steenbeke; Matilda Halloran; John Fanton; Michael K. Axthelm; Norman L. Letvin; Joseph Sodroski

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Michael Farzan

Scripps Research Institute

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