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Stimulation of camel polyclonal antibody against human T cell
immunoglobulin and Mucin-3
Homayouni,V.
Isfahan University of Medical Sciences, Isfahan, Iran, Islamic Republic of
Background:
T cell Immunoglobulin and Mucin (TIM)-3 is a type I
transmembrane glycoprotein and amember of theTIM family which
is identified as a receptor. This receptor expresses on T helper type 1
(Th1) cells and binds to galectin-9 (Gal9). This interaction induces an
inhibitory signal, resulting inapoptosisofTh1cells andcytotoxicCD8
T cells in vitro. Antibody therapy for immune checkpoint blockade
has achieved promising results for several types of malignant
tumors. Therefore, this immunomodulatory molecule may be used
as a novel target for clinical purposes.The productionof camel heavy
chain antibodies (HCAbs) against TIM-3-expressing cell line was
reported in this study.
Methods:
A pre-synthesized human TIM-3cDNA was insertd into
pcDNA3.1 plasmid and the new construct was transfected in HEK
cell. TIM-3 expressionwas confirmed by qRT-PCR and flowcytometry
methods. A 6 months old camel was immunized with the lysate
prepared from rTIM-3 expressing HEK cells 4times.Then, the anti-
TIM-3 antibody level was evaluated using ELISA method.
Results and conclusion:
TIM-3 was successfully cloned in HEK cells
and more than 88% of the cells expressed TIM-3. Hence, using HEK
cells was produced a readily obtainable source of TIM-3. It was used
for camel immunization. High level of anti-TIM-3 antibody was
detected in its serum after the final injection. This antibody may be
useful for future diagnostic or therapeutic approaches.
B cell depletion compromises CD8+ T cell response in murine
T
cruzi
infection
FioccaVernengo,F.,Beccaria,C.G.,AraujoFurlán,C.,ToselloBoari,J.,
GorositoSerrán,M.,Montes,C.,AcostaRodriguez, E.V., Gruppi,A.
SchoolofChemistry.NationalUniversityofCórdoba,Córdoba,
Argentina
Considering CD8+T cells play a major role in
T. cruzi
protective
immunity, the signals that promote the generation and
maintenance of CD8+T cell responses needs to be identified. Many
reports highlight the role of B cells in promoting cellular immunity;
however, if B cells affect CD8 responses remain uncharacterized.
To assess B-cell function in promoting CD8 responses in Chagas
disease, C57BL/6 mice were intraperitoneally injected with anti-
CD20, to deplete B cells, or with control antibodies. Eight days after
treatment, mice were infected with 5000 trypomastigotes. Flow
cytometric analysis using tetramer staining revealed that at day 14
post infection, depleted mice had a lower frequency of parasite-
specific CD8+T cells in spleen, blood and liver compared to control
mice (p=0,0003, 0,0353 and 0,0447 respectively). Interestingly, B-cell
deficient mice showed higher percentages of naïve CD8+T cells
(CD44-CD62L+)andlowerfrequencyofeffectormemoryCD8+Tcells
(CD44+CD62L-). CD8+T cells from B-cell depleted mice presented
a Memory Precursor Effectors Cells phenotype unlike CD8+T cells
from B-cell sufficient mice who exhibit a Short Lived Effectors
Cells phenotype. Also, we found that depleted mice had lower
frequencies of IFN-γ- or TNF-producing CD8+T cells and displayed
lower apoptosis and proliferation rate. Unexpectedly CD8+T cells
from B-cell depleted mice overexpressed molecules associated
to activation/migration than control mice. These results identify
functional defects in CD8+T cells generated in B cell-depleted mice,
highlighting B cell influence on CD8+T cell response. Furthermore,
our study urges caution during B-cell depletion therapies due to its
possible side effects on the immunity against infections.
Transgenic expression of N-acetylglucosaminyltransferase V
in T cells accelerates autoimmune diabetes in NOD mice by
enhancing pathogenicity of CD8 T cells
Chien,M.-W.
1
,Khoo,K.-H.
2
,Sytwu,H.-K.
1
1
National Defense Medical Center, Department and Graduate
Institute of Microbiology and Immunology, Taipei, Taiwan, Republic
of China,
2
Academia Sinica, Institute of Biological Chemistry, Taipei,
Taiwan, Republic of China
β1,6
N-acetylglucosaminyltransferase
V
(Mgat5)
is
a
glycosyltransferasethatincreasesN-glycanbranchingbytransferring
N- acetylglucosamine (GlcNAc) fromUDP-GlcNAc to hydroxyl group
on carbon 6 of the α1,6-linked mannose of N-glycans. Previous
studies have shown that deficiency of Mgat5 in mice decreased
the T cell activation thresholds and exacerbated the incidence of
experimental autoimmune encephalomyelitis (EAE). The non-obese
diabetic (NOD) mice, a widely used animal model for human type 1
diabetes (T1D), developed spontaneous autoimmune diabetes and
mainly caused byT cell-dependent destruction of pancreatic β-cells.
Recently,NODmouseisreportedasanEAEsusceptiblestrainsinceless
N-glycan branching expressed on T cells. However, the relationship
between Mgat5-mediated N-glycan branching and diabetogenic
processingonT cells inNOD mice is still unknown. Here, weobserved
that Mgat5 transgenic T cells expressed higher level of N-glycan
branching and had attenuated proliferation potential compared to
control littermate T cells in NOD mice. Unexpectedly, a significantly
higher diabetic incidence was found in both Mgat5 transgenic
NOD mice and Mgat5/NY8.3 doubly transgenic mice that express a
highly pathogenic transgenic TCR on CD8 T cells. Further, the higher
diabetic incidence was also found in NOD/SCID recipients received
adoptive transfer of Mgat5-overexpressed CD8 T cells, indicating
that Mgat5 overexpression enhances the pathogenic properties of
CD8T cells in an antigen-specificmanner. Taken together, our results
suggest that upregulated Mgat5-mediated N- glycan branching on
CD8T cells contributes to autoimmune diabetes inNODmice.
Features of phenotypic structure and functional activity of
cord blood immune cells depending a gestational age
Lyapunov,V.
1
,Chistyakova,G.
1
,Bychkova,S.
2
,Remizova,I.
1
,Chereshnev,
V.
3
,Ustyantseva,L.
2
1
MotherandChildCareResearhInstituteinRussianPublicHealth
Ministry,ImmunologyandClinicalMicrobiology,Ekaterinburg,
RussianFederation,
2
MotherandChildCareResearhInstituteinRussian
PublicHealthMinistry,PathologyofPrematureInfants, Ekaterinburg,
RussianFederation,
3
InstituteofImmunologyandPhysiologyUBRAS,
Ekaterinburg,RussianFederation
In order to identify genotypic characteristics of the composition and
functional activity of immune cells of umbilical cord blood (UCB) as a
function of gestational age (GA), studied 102 UCB samples of babies
born at 25-28, 29-32 and 33-36 weeks of GA and 31 UCB samples
from comparison group (37-41 weeks GA). By the method of flow