Analysis of the expressed heavy chain variable-region genes of Macaca fascicularis and isolation of monoclonal antibodies specific for the Ebola virus' soluble glycoprotein

Chris Druar, Surinder S. Saini, Meredith A. Cossitt, Fei Yu, Xiangguo Qiu, Thomas Geisbert, Steven Jones, Peter B. Jahrling, Donald I H Stewart, Erik J. Wiersma

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The cynomolgus macaque, Macaca fascicularis, is frequently used in immunological and other biomedical research as a model for man; understanding it's antibody repertoire is, therefore, of fundamental interest. The expressed variable-region gene repertoire of a single M. fascicularis, which was immune to the Ebola virus, was studied. Using 5′ rapid amplification of cDNA ends with immunoglobulin (Ig)G-specific primers, we obtained 30 clones encoding full-length variable, diversity, and joining domains. Similar to the human VH repertoire, the M. fascicularis repertoire utilized numerous immunoglobulin heavy variable (IGHV) gene fragments, with the VH3 (41%), VH4 (39%), and VH1 (14%) subgroups used more frequently than the VH5 (3.9%) or VH7 (1.7%) subgroups. Diverse immunoglobulin heavy joining (IGHJ) fragments also appeared to be utilized, including a putative homolog of JH5β gene segment identified in the related species Macaca mulatta, Rhesus macaque, but not in humans. Although the diverse V region genes in the IgG antibody repertoire of M. fascicularis had likely undergone somatic hypermutations (SHMs), they nevertheless showed high nucleotide identity with the corresponding human germline genes, 80-89% for IGHV and 72-92% for IGHJ. M. fascicularis and human VH genes were also similar in other aspects: length of complementarity-determining regions and framework regions, and distribution of consensus sites for SHMs. Finally, we demonstrated that monoclonal antibodies (mAbs) specific for an Ebola protein could be obtained from M. fascicularis tissue samples by phage display technology. In summary, the study provides new insight into the M. fascicularis V region gene repertoire and further supports the idea that macaque-derived mAbs may be of therapeutic value to humans.

Original languageEnglish (US)
Pages (from-to)730-738
Number of pages9
JournalImmunogenetics
Volume57
Issue number10
DOIs
StatePublished - Nov 2005
Externally publishedYes

Fingerprint

Ebolavirus
Macaca fascicularis
Glycoproteins
Monoclonal Antibodies
Immunoglobulins
Genes
Macaca
Macaca mulatta
Immunoglobulin G
Complementarity Determining Regions
Antibodies
Bacteriophages
Biomedical Research
Nucleotides
Complementary DNA
Clone Cells
Technology

Keywords

  • Antibody repertoire
  • Cynomolgus monkey
  • Ebola virus
  • Monoclonal antibody
  • V gene

ASJC Scopus subject areas

  • Immunology
  • Genetics

Cite this

Analysis of the expressed heavy chain variable-region genes of Macaca fascicularis and isolation of monoclonal antibodies specific for the Ebola virus' soluble glycoprotein. / Druar, Chris; Saini, Surinder S.; Cossitt, Meredith A.; Yu, Fei; Qiu, Xiangguo; Geisbert, Thomas; Jones, Steven; Jahrling, Peter B.; Stewart, Donald I H; Wiersma, Erik J.

In: Immunogenetics, Vol. 57, No. 10, 11.2005, p. 730-738.

Research output: Contribution to journalArticle

Druar, Chris ; Saini, Surinder S. ; Cossitt, Meredith A. ; Yu, Fei ; Qiu, Xiangguo ; Geisbert, Thomas ; Jones, Steven ; Jahrling, Peter B. ; Stewart, Donald I H ; Wiersma, Erik J. / Analysis of the expressed heavy chain variable-region genes of Macaca fascicularis and isolation of monoclonal antibodies specific for the Ebola virus' soluble glycoprotein. In: Immunogenetics. 2005 ; Vol. 57, No. 10. pp. 730-738.
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abstract = "The cynomolgus macaque, Macaca fascicularis, is frequently used in immunological and other biomedical research as a model for man; understanding it's antibody repertoire is, therefore, of fundamental interest. The expressed variable-region gene repertoire of a single M. fascicularis, which was immune to the Ebola virus, was studied. Using 5′ rapid amplification of cDNA ends with immunoglobulin (Ig)G-specific primers, we obtained 30 clones encoding full-length variable, diversity, and joining domains. Similar to the human VH repertoire, the M. fascicularis repertoire utilized numerous immunoglobulin heavy variable (IGHV) gene fragments, with the VH3 (41{\%}), VH4 (39{\%}), and VH1 (14{\%}) subgroups used more frequently than the VH5 (3.9{\%}) or VH7 (1.7{\%}) subgroups. Diverse immunoglobulin heavy joining (IGHJ) fragments also appeared to be utilized, including a putative homolog of JH5β gene segment identified in the related species Macaca mulatta, Rhesus macaque, but not in humans. Although the diverse V region genes in the IgG antibody repertoire of M. fascicularis had likely undergone somatic hypermutations (SHMs), they nevertheless showed high nucleotide identity with the corresponding human germline genes, 80-89{\%} for IGHV and 72-92{\%} for IGHJ. M. fascicularis and human VH genes were also similar in other aspects: length of complementarity-determining regions and framework regions, and distribution of consensus sites for SHMs. Finally, we demonstrated that monoclonal antibodies (mAbs) specific for an Ebola protein could be obtained from M. fascicularis tissue samples by phage display technology. In summary, the study provides new insight into the M. fascicularis V region gene repertoire and further supports the idea that macaque-derived mAbs may be of therapeutic value to humans.",
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