9 / 25
Page 71
Notes:
Journal of Analytical & Bioanalytical Techniques | ISSN: 2155-9872 | Volume 9
World HPLC, Separation Techniques & Pharmacovigilance
World Analytical Chemistry & Mass Spectrometry
18
th
International Conference on
August 29-30, 2018 | Toronto, Canada
&
Comparison of immune cells subsets,
ex-vivo
and
in-vivo
expression of T cell activation and memory
marker between LNC and corresponding PBMC from Calves Exposed to Natural
Mycobacterium
bovis
Infection
Fekadu Desta
1, 3, 4
, Gobena Ameni
1
, F Javier Salguero Bodes
2
and
awleigh Howe
3
1
Addis Ababa University, Ethiopia
2
University of Surrey, Guildford, UK
3
Armaure Hansen Research Institute, Ethiopia
4
Wollo University, Ethiopia
C
ell-mediated immunity and development of necrotic granulomas in
Mycobacterium bovis (M. bovis)
infected lymph node
(LN) is pathognomonic for bovine tuberculosis (BTB). This delayed hypersensitive host response involves a complex
interaction of cellular and immune mediators within systemic circulation and LN. Hence, tuberculosis immunological
response should be independently investigated at the peripheral blood and LN tissue level. The objective of this study was,
therefore, to compare the cell surface and cytokine expression between immune cell from peripheral blood and lymph node
cells (LNC) from calves on BCG efficacy trial. Twenty pairs of peripheral blood mononuclear cells (PBMC) and) LNC fromM.
bovis naturally infected calves during BCG vaccine experiment trial were isolated and investigated in two phases of the flow-
cytometry experiment. In the first phase of a flow-cytometry experiment the proportion of
ex-vivo
CD25+ expressing cells was
significantly higher (P<0.05) in CD4+ and CD8+ T node than that of peripheral blood. However, such difference in CD25+
expression was not observed in WC1 γδ T cells. Contrary to CD25+
ex-vivo
expression,
in-vitro
IFN-γ and TNF-α producing
cells were greater (P<0.05) in T cells of the peripheral blood than T cells of lymph node after PMA + ionomycin stimulation.
This difference in IFNγ and TNFα responses was also statistically significant between a vaccinated and non-vaccinated group.
An IL-4 producing cell was not evident in PBMC and LNC. During the second phase of flow-cytometry experiment additional
surface marker; CD2, CD21, CD205, CD335 and CD1W2 were included to add more panels for immune cell subset. The
second experiment revealed that PBMC CD4–WC1+ and CD8–WC1+ γδ T cells and CD205+D1W2+ DC subset exhibited
lower percentage than γδ T cells and DC of LNC respectively (p=0.0001, p=0.0061). However, PBMC CD335+CD2+ NKT
cells subset exhibited a higher percentage than NKT cells of LNC (p=0.0129). No difference was observed between groups
in the percentage of the rest of T-cell and B-cell (p>0.05). Findings of this study suggest the existence of phenotypic immune
compartmentalization between the two tissue compartments.
Biography
Fekadu Desta is a veterinarian with a dream and responsibility to control and prevent the existing endemic and emerging new diseases of livestock and companion
animals, Ethiopia. For the last 6 years, he has been doing his PhD in Tropical Infectious Diseases with a PhD dissertation entitled “Comparison of Immune
Cell Subsets,
ex-Vivo
and
in-Vitro
Expression of Activation and Memory Marker Between LNC and the Corresponding PBMC from Calves Exposed to Natural
Mycobacterium bovis Infection in BCG Efficacy Trial” which is one of a research priority of the country. Currently, he is on data analysis, interpretation and result
dissemination stage. He has published one paper on molecular epidemiology of M. bovis and preparing 3 more manuscript on bovine immunology.
fekadudg@gmail.comFekadu Desta et al., J Anal Bioanal Tech 2018, Volume 9
DOI: 10.4172/2155-9872-C1-028