Immune responses to human factor IX in haemophilia B mice of different genetic backgrounds are distinct and modified by TLR4

Brandon Wilder, X. Wang, A. Sherman, G. L. Rogers, D. M. Markusic

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Our laboratory develops protocols to prevent or reverse ongoing anti-hFIX IgG inhibitors in haemophilia B mice with a F9 gene deletion on BALB/c and C3H/HeJ backgrounds. C3H/HeJ F9-/Y mice develop high titre anti-hFIX IgG1 inhibitors and anaphylaxis, whereas most BALB/c F9-/Y mice have mild anti-hFIX IgG1 inhibitors and no anaphylaxis. Our aim was to determine if hFIX-specific B- and T-cell responses in BALB/c and C3H/HeJ F9-/Y mice trigger the difference in anti-hFIX immune responses. BALB/c and C3H/HeJ F9-/Y mice were challenged weekly with recombinant hFIX protein. Humoral immune responses were determined by IgG1 and IgG2a anti-hFIX ELISA, Bethesda assay for inhibitors and B-cell ELISpot on bone marrow and spleen cells. T-cell studies measured the TH1 (IFN-γ) and TH2 (IL-4) cytokine responses in splenocytes at the mRNA and protein level in response to hFIX protein. Antibody responses were also measured in C3H/HeJ/OuJ F9-/Y mice with restored toll-like receptor 4 (TLR4) function. BALB/c F9-/Y mice have a TH2 skewed response and a reduction in anti-hFIX secreting plasma cells in the bone marrow. Independent antigen challenge revealed both strains generated equivalent IgG1 antibody titres to an intravenously delivered antigen. C3H/HeJ F9-/Y mice have a mixed TH1 and TH2 response (mainly TH2). Importantly, TLR4 signalling has a modulatory role in the C3H background on the levels of anti-hFIX IgG1 and incidence of anaphylaxis. The background strain strongly impacts the immune response to hFIX, which can be significantly impacted by mutations in innate immune sensors.

Original languageEnglish (US)
Pages (from-to)133-139
Number of pages7
JournalHaemophilia
Volume21
Issue number1
DOIs
StatePublished - Jan 1 2015
Externally publishedYes

Fingerprint

Hemophilia B
Toll-Like Receptor 4
Factor IX
Immunoglobulin G
Anaphylaxis
T-Lymphocytes
Antigens
Genetic Background
Gene Deletion
Humoral Immunity
Plasma Cells
Recombinant Proteins
Bone Marrow Cells
Interleukin-4
Antibody Formation
Proteins
B-Lymphocytes
Spleen
Bone Marrow
Enzyme-Linked Immunosorbent Assay

Keywords

  • Anaphylaxis
  • BALBc
  • C3H/HeJ
  • Factor IX
  • Haemophilia B murine model
  • Immune response

ASJC Scopus subject areas

  • Hematology
  • Genetics(clinical)

Cite this

Immune responses to human factor IX in haemophilia B mice of different genetic backgrounds are distinct and modified by TLR4. / Wilder, Brandon; Wang, X.; Sherman, A.; Rogers, G. L.; Markusic, D. M.

In: Haemophilia, Vol. 21, No. 1, 01.01.2015, p. 133-139.

Research output: Contribution to journalArticle

Wilder, Brandon ; Wang, X. ; Sherman, A. ; Rogers, G. L. ; Markusic, D. M. / Immune responses to human factor IX in haemophilia B mice of different genetic backgrounds are distinct and modified by TLR4. In: Haemophilia. 2015 ; Vol. 21, No. 1. pp. 133-139.
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abstract = "Our laboratory develops protocols to prevent or reverse ongoing anti-hFIX IgG inhibitors in haemophilia B mice with a F9 gene deletion on BALB/c and C3H/HeJ backgrounds. C3H/HeJ F9-/Y mice develop high titre anti-hFIX IgG1 inhibitors and anaphylaxis, whereas most BALB/c F9-/Y mice have mild anti-hFIX IgG1 inhibitors and no anaphylaxis. Our aim was to determine if hFIX-specific B- and T-cell responses in BALB/c and C3H/HeJ F9-/Y mice trigger the difference in anti-hFIX immune responses. BALB/c and C3H/HeJ F9-/Y mice were challenged weekly with recombinant hFIX protein. Humoral immune responses were determined by IgG1 and IgG2a anti-hFIX ELISA, Bethesda assay for inhibitors and B-cell ELISpot on bone marrow and spleen cells. T-cell studies measured the TH1 (IFN-γ) and TH2 (IL-4) cytokine responses in splenocytes at the mRNA and protein level in response to hFIX protein. Antibody responses were also measured in C3H/HeJ/OuJ F9-/Y mice with restored toll-like receptor 4 (TLR4) function. BALB/c F9-/Y mice have a TH2 skewed response and a reduction in anti-hFIX secreting plasma cells in the bone marrow. Independent antigen challenge revealed both strains generated equivalent IgG1 antibody titres to an intravenously delivered antigen. C3H/HeJ F9-/Y mice have a mixed TH1 and TH2 response (mainly TH2). Importantly, TLR4 signalling has a modulatory role in the C3H background on the levels of anti-hFIX IgG1 and incidence of anaphylaxis. The background strain strongly impacts the immune response to hFIX, which can be significantly impacted by mutations in innate immune sensors.",
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