Document 0213
 DOCN  M9440213
 TI    Neutralization sensitivity of human immunodeficiency virus type 1 is
       determined in part by the cell in which the virus is propagated.
 DT    9404
 AU    Sawyer LS; Wrin MT; Crawford-Miksza L; Potts B; Wu Y; Weber PA; Alfonso
       RD; Hanson CV; Viral and Rickettsial Disease Laboratory, California
       Department; of Health Services, Berkeley 94704.
 SO    J Virol. 1994 Mar;68(3):1342-9. Unique Identifier : AIDSLINE
       MED/94149821
 AB    Neutralizing antibody responses to human immunodeficiency virus type 1
       (HIV-1) vary widely and have not been reproducibly associated with
       prognosis or disease progression. We have found that both low-passage
       clinical isolates and laboratory-adapted strains of HIV-1 have different
       sensitivities to neutralization by the same antiserum, depending on the
       host cell in which the viral stock is prepared. One such isolate (VL069)
       grown in H9 cells was neutralized by 20 human sera at a geometric mean
       titer of 1:2,047; this same isolate prepared in peripheral blood
       mononuclear cell (PBMC) culture was neutralized at a mean titer of <
       1:10 by the same sera. Adsorption and mixing experiments indicated that
       neither antibody to H9 cell components nor blocking by excess viral
       antigen was responsible for the differences observed. This host cell
       effect is rapidly reversible upon passage of the virus from PBMCs to H9
       cells and back into PBMCs. In contrast, the neutralization
       characteristics remained remarkably stable over extended culture in
       PBMCs. Two laboratory strains and five clinical isolates were evaluated
       in expanded studies of this phenomenon. While the neutralization
       characteristics of most of the strains studied were affected by the host
       cell in which the strain was propagated, two of the strains (one
       clinical isolate and one laboratory strain) appeared antigenically
       unaffected by their cell of origin. Host cell effect was also evident in
       neutralization by monoclonal antibodies directed against the CD4-binding
       region and the V2, V3, and gp41 regions. Possible mechanisms for this
       host cell effect include (i) mutation during passaging; (ii) selection
       in different host cells of different subpopulations of the (uncloned)
       viral stock; and (iii) cell-specific posttranslational modifications. To
       explore these possibilities, the V3 through V5 region of gp120 was
       sequenced in preparations made by passing VL069 into H9 cells and into
       PBMCs; HIVMN grown in CEM-SS cells and in PBMCs was also sequenced. In
       both cases, a few amino acid changes outside the V3 region were found.
       Studies are currently under way to assess the significance of these
       changes.
 DE    Adaptation, Biological  Amino Acid Sequence  Antibodies, Monoclonal
       Cells, Cultured  Comparative Study  Human  HIV Antibodies/*IMMUNOLOGY
       HIV Envelope Protein gp120/GENETICS/IMMUNOLOGY  HIV Envelope Protein
       gp41/GENETICS/IMMUNOLOGY  HIV-1/*GROWTH & DEVELOPMENT/*IMMUNOLOGY
       Molecular Sequence Data  *Neutralization Tests  Serial Passage  Species
       Specificity  Virus Cultivation  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).