Effects of CHO-expressed recombinant lactoferrins on mouse dendritic cell presentation and function

Abstract

Lactoferrin (LF), a natural iron-binding protein, has previously demonstrated effectiveness in enhancing the Bacillus Calmette–Guérin (BCG) tuberculosis vaccine. This report investigates immune modulatory effects of Chinese hamster ovary (CHO) cell-expressed recombinant mouse and human LFs on mouse bone marrow-derived dendritic cells (BMDCs), comparing homologous and heterologous functions. BCG-infected BMDCs were cultured with LF, and examined for class II presentation molecule expression. Culturing of BCG-infected BMDCs with either LF decreased the class II molecule-expressing population. Mouse LF significantly increased the production of IL-12p40, IL-1β and IL-10, while human LF-treated BMDCs increased only IL-1β and IL-10. Overlaying naïve CD4 T-cells onto BCG-infected BMDCs cultured with mouse LF increased IFN-γ, whereas the human LF-exposed group increased IFN-γ and IL-17 from CD4 T cells. Overlay of naïve CD8 T cells onto BCG-infected BMDCs treated with mouse LF increased the production of IFN-γ and IL-17, while similar experiments using human LF only increased IL-17. This report is the first to examine mouse and human recombinant LFs in parallel experiments to assess murine DC function. These results detail the efficacy of the human LF counterpart used in a heterologous system to understand LF-mediated events that confer BCG efficacy against Mycobacterium tuberculosis challenge.

Recombinant human lactoferrin modulates human PBMC derived macrophage responses to BCG and LPS

“…This activity of lactoferrin has been well documented with bovine, recombinant human and recombinant mouse lactoferrins on mouse bone marrow derived macrophages and dendritic cells. Co-culture experiments demonstrated that CD86 high expressing cells do increase T cell activation [11,12,51,52]. Additionally, this increase in CD86 has been correlated with in vivo data that demonstrated mice immunized with BCG and lactoferrin (bovine and human) enhanced memory recall responses that led to enhanced pathological protection against MTB challenge [13–17,20,53].…”

Lactoferrin: A Modulator for Immunity against Tuberculosis Related Granulomatous Pathology

“…Preliminary investigations show that a newly developed, recombinant mouse lactoferrin produced in CHO stable cell lines functions in a manner very similar to the bovine and human recombinant, to limit immunopathology after oral delivery. Mechanistically, it was shown that the mouse and human forms are nearly identical in modulation of dendritic cell function in response to Bacillus Calmette-Guerin (BCG) [ 25 , 109 ]. The dendritic cell population responds with increased inflammatory cytokines and a shift towards MHC Class II expression when BCG is combined with the lactoferrins.…”

Oral recombinant human or mouse lactoferrin reducesMycobacterium tuberculosisTDM induced granulomatous lung pathology

“…Recently, both recombinant human and mouse lactoferrins were produced in sufficient quantities in the Chinese hamster ovary (CHO) cell line. Previous studies published on mouse-derived immune cells on CHO-derived recombinant human and mouse lactoferrins demonstrated similar immune modulatory effects (Hwang et al 2015; O’Shea et al 2015), suggesting that the mouse model can be used as a preclinical model for evaluating efficacy of recombinant human lactoferrin.…”