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.com
Volume 7, Issue 2 (Suppl)
J Ecosyst Ecography, an open access journal
ISSN:2157-7625
September 18-20, 2017
September 18-20, 2017 Toronto, Canada
Joint Conference
International Conference on
International Conference on
Environmental Microbiology and Microbial Ecology
&
Ecology and Ecosystems
Monoclonal and polyclonal antibodies directed toward Tsukamurella pulmonis polysaccharide
Adnan Saeed, Mariola Paściak, Jerzy Szkudlarek, Marek Drab
and
Andrzej Gamian
Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Poland
T
he genus
Tsukamurella
includes aerobic Gram-positive and modified acid alcohol-fast-positive rods belonging to the aerobic
actinomycetes, usually found in soil, sludge and arthropods.
T. pulmonis
is a rare human pathogen associated with oncologic
and immunosuppressed patients, and a variety of infections have been associated with this bacterium: pneumonia, conjunctivitis,
keratitis and catheter-related bacteremia. The strain of
T. pulmonis
was obtained from Polish Collection of Microorganisms (PCM
2578). The bacteria were identified morphologically by Gram staining, scanning electron microscopy and MALDI-TOF mass
spectrometry. The polysaccharides were extracted by trichloroacetic acid from dry bacterial cell mass and purified by anion exchange
and gel permeation chromatography. Sugar composition was determined by gas-liquid chromatography-mass spectrometry (GLC-
MS). The monoclonal antibodies against polysaccharide of
T. pulmonis
were obtained by the hybridoma technique. Rabbit polyclonal
immune sera against
T. pulmonis
were obtained by immunization with
T. pulmonis
whole cells. The scanning electron microscopy
showed a variation of the rod shape of the bacteria. Purified polysaccharide of
T. pulmonis
consists of arabinose and mannose also
traces amounts of glucose and galactoses were detected. Two hybridomas 5 and 23 producing mAbs against polysaccharide antigen
were IgM class. The ELISA test allowed to detect cross reactivity of these monoclonal antibodies with some other Actinomyces spp
antigens. Reactivity in double immunodiffusion test of polysaccharide antigen of
T. pulmonis
with polyclonal sera against
T. pulmonis,
T. tyrosinosolvens
,
T. inchonensis
and
T. paurometabola
cells was observed. Cross reactivity of EPS were seen with all Tsukamurella
antisera studied. Results indicate that monoclonal and polyclonal antibodies and polysaccharide could serve as tools for diagnostic
purposes but their diagnostic potential should be further studied regarding the specificity and structure of epitope recognized.
Biography
Adnan Saeed is researcher in Department of Immunology and Infectious Diseases, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish
Academy of Sciences, Weigla 12, Wroclaw, Poland.
Adnan.m14.m14@gmail.comAdnan Saeed et al., J Ecosyst Ecography 2017, 7:2 (Suppl)
DOI: 10.4172/2157-7625-C1-029