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Cellular expression and antimicrobial function of a phylogenetically conserved novel histone 1x‐like protein
Author(s) -
JasoFriedmann Liliana,
Connor Meghan A.,
Moss Lauren D.,
Lackay Sarah N.,
Leary John H.,
Evans Donald L.
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.672.42
Subject(s) - polyclonal antibodies , monoclonal antibody , microbiology and biotechnology , extracellular , biology , chemistry , antibody , immunology
Soluble linker and core histone‐like proteins isolated from many different types of mucosal and lymphoreticular tissues have antimicrobial activity. Previous reports by others have shown that granule extracts from RAW 264.7 cells and neutrophil extracellular traps contain H1 that may function in bacterial killing. We have previously identified an H1x‐like protein (i.e. NCC cationic antimicrobial protein‐s/ncamp‐1) from fish NK‐like cells (nonspecific cytotoxic cells/NCC) and prepared polyclonal and monoclonal antibodies to this membrane protein. In the present study we identify the (ICR) mouse ncamp‐1 equivalent using polyclonal as well as an anti‐ncamp‐1 monoclonal antibody (mab 9C9). Polychromatic flow cytometry revealed mab 9C9 expression on PBL (Gr‐1 + 21.1% ncamp‐1; DX‐5 + 12.2% + ); mesenteric lymph node (CD11c = 42.3% ncamp‐1, Gr‐1 = 24.6%; CD21 = 33.3%); and splenocytes had approximately the same distribution and percentages of ncamp‐1 + cells as the LN. Granule extracts were prepared from RAW 264.7 cells and analyzed by Western blots. A 29‐32 kDa signal was produced. Granule extracts were also prepared for an antimicrobial assay. A significant reduction in E. coli CFU was produced by a RAW 264.7 granule extract. The specificity of bacterial killing by this extract was determined by pretreatment of the extract with polyclonal anti‐ncamp‐1. This produced a significant reduction in killing. Pretreatment with pre‐immune serum did not reduce bacterial cell killing by the extract. These studies provided preliminary data that a potential novel class of pattern recognition receptor is phylogenetically conserved and exists on mouse lymphoreticular cells and in macrophage granules. Research supported by USDA award # 2006‐35204‐16562.