(A) The frequency of anti-nuclear new emigrant B cells in IRAK-4 and
MyD88-deficient patients is higher than in controls and UNC-93B-deficient
patients. Each diamond represents an individual, and the average is shown
with a bar.
(B) The frequencies of chromatin non-reactive and reactive clones in
IRAK-4- and MyD88-deficient patients are higher than in controls and
UNC-93B-deficient patients. Histograms represent means ± s.e.m.
of frequencies of chromatin non-reactive (open bars) or chromatin reactive
(black bars) antibodies. Statistically significant differences between
patients and controls are indicated.
(C) ANAs from IRAK-4- and MyD88-deficient new emigrant B cells show
various patterns of anti-nuclear staining. White arrows indicate dividing
cells that demonstrate negative (top row) or positive (bottom row) staining
for the condensed chromatin material.
(D) Increased anti-kinetoplast antibodies in new emigrant B cells
from IRAK-4- (black bars) and MyD88-deficient (dark grey bars) compared to
controls (open bars) and UNC-93B-deficient patients (light grey bars).
Histograms represent means ± s.e.m. of proportion of kinetoplast
reactive clones. Statistically significant differences between patients and
controls are indicated.
(E) IRAK-4- and MyD88-deficient clones express ANAs that can
recognize the kinetoplast of C. Luciliae. White and red
arrows indicate positively stained kinetoplasts and nucleus of C.
Luciliae respectively.
(F) Kinetoplast reactive ANA-expressing B cells display the highest
frequency of positively charged residues in their heavy chain CDR3s. New
emigrant B cells from IRAK-4 and MyD88-deficient patients were grouped into
non-autoreactive, autoreactive-non ANA (HEp-2-reactive and/or polyreactive),
kinetoplast non-reactive ANA and kinetoplast reactive ANA clones. Pie charts
show the proportion of heavy chain CDR3s with 0, 1, 2 and 3 or more positive
charges. Absolute numbers of clones analyzed in each B cell fraction is
indicated in the center. The average number of positively charged residues
per CDR3 is indicated below each fraction. The numbers
of positively charged residues in immunoglobulin heavy chain CDR3s were
calculated excluding the arginin or lysin found at position 94 in most germ
line VHs.