Fig. 1: Reactivity of recombinant IgD+ LP cell-derived Fabs with bacterial lysates. | Nature Communications

Fig. 1: Reactivity of recombinant IgD+ LP cell-derived Fabs with bacterial lysates.

From: Lymphocyte predominant cells detect Moraxella catarrhalis-derived antigens in nodular lymphocyte-predominant Hodgkin lymphoma

Fig. 1

a Representative Immuno-dot blots of bacterial lysates. NLPHL-derived recombinant Fabs of six cases bound to lysates of M. catarrhalis, and of one case to lysate of M. osloensis. Eight cases showed no reactivity against bacterial lysates. All six M. catarrhalis-reactive recombinant Fabs (#3, #6, #9, #10, #13, and #14) and the one M. osloensis-reactive Fab (#11) were derived from IgD+ NLPHL cases. b Representative Western blot of recombinant C-terminally FLAG tagged M. catarrhalis RpoC expressed in HEK293 cells confirmed reactivity of lymphoma Fabs #3, #6, #9, #10, #13, and #14 with the recombinant bacterial antigen. A recombinant C-terminally FLAG-tagged SMCHD1 fragment was used as negative control. c ELISA with recombinant C-terminally FLAG-tagged M. catarrhalis RpoC as a coat demonstrated specific reactivity of Fabs derived from LP cells of six patients (#3, #6, #9, #10, #13, and #14) against recombinant RpoC. Values are mean ± SD. d Determination of the epitope of RpoC recognized by IgD+ NLPHL Fabs. Recombinant Fabs #3, #6, #9, and #10 bind AA 600–1130 of M. catarrhalis RpoC. Values are mean ± SEM. e Serum reactivity against RpoC in patients from whom recombinant NLPHL Fabs were derived. The two IgD+ NLPHL patients with RpoC-reactive recombinant lymphoma Fabs (#3 and #6) had higher anti-RpoC-antibody serum titers (1:1600) than a healthy control with anti-RpoC-antibodies (1:200). The IgD+ NLPHL patient with the M. osloensis-specific Fab (#11) had no serum anti-RpoC-antibodies. Values are mean ± SD. Experiments were independently repeated three times. f: Immunoglobulin classes and subclasses and light chains of serum RpoC antibodies from the two patients with RpoC-reactive BCR of the LP cells (#3 and #6) and a seropositive healthy control. Patients #3 and #6 both show a predominance of IgG1/κ. The healthy control had additionally anti-RpoC-antibodies of IgM class and in contrast to patients #3 and #6 no clear light chain restriction. Values are mean ± SD. g Representative Western blot of FLAG-tagged M. osloensis SUCLG1 expressed in HEK293 cells confirms reactivity of lymphoma Fab #11 with the recombinant bacterial antigen. h ELSIA with recombinant M. osloensis SUCLG1 as a coat demonstrated specific reactivity of Fabs derived from LP cells of patient #11 against recombinant SUCLG1. Values are mean ± SD. Data in a–c, e, and f are representative of three independent experiments each; data in d, g, and h are representative of two independent experiments.

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