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Register a new userComparative Study For Promastigote & Amastigote Stages Antigens of Cutaneous Leishmania Tropica
دراسة مقارنة لمستضدات طوري الليشمانيا الجلدية المدارية أمامي السوط Promastigote و عديم السوط Amastigote
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Damascus University ورقة بحثية
Publication date
2011
fields
Biology
and research's language is
العربية
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Shamra Editor
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ترمي هذه الدراسة إلى الوقوف على المحتوى البروتيني للطورين الرئيسين لليشمانيا: أمامي السوط promastigote و عديم السوط amastigote و لتعيين الأوزان الجزيئية للبروتينات الأكثر تعبيراً و ذلـك بوساطة تقنية الرحلان الكهربائي العمودي و من ثم تحديد أهم مستضدات هذين الطورين بتقنية التبـصيم المناعي و باستخدام أمصال فئران منعت بالخلاصات البروتينية لهذه الأطوار و مقارنتها ببعـضها بهـدف تحديد المستضدات المشتركة بينهما و المستضدات الخاصة بكل طور، و ذلك بغية اسـتخدامها فـي إطـار استراتيجية تلقيحية تجاه الليشمانيا.
The parasitic antigens are of great interest in immune responses and in the strategies for developing vaccines. The aim of this study was to identify the major antigens which are expressed in the two leishmania tropica stages, promastigote & amastigote, employing the western blot analysis using the sera collected from mice immunized with the above mentioned parasite stages' total protein.
References used
Beetham J. K, Donelson J. E, and Dahlin R. R. (2003). Surface glycoprotein PSA (GP46) expression during short-and long-term culture of leishmania chagasi. Mol Biochem Parasitol 131(2): 109-117
Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72:248-254
Champsi J and McMahon-Pratt. (1988). Membrane glycoprotein M-2 protects against leishmania amazonensis infection. Infect. Immun.56:3272- 3279
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Kinetoplastid membrane protein-11 (KMP-11), a protein present in all kinetoplastid
protozoa studied up to date, is considered a potential vaccine candidate for Leishmaniasis.
Such vaccine molecules must be expressed in amastigotes which represent t
he infective
forms for mammals, while promastigotes are the flagellate forms found in the insect hosts.
However, the expression of KMP-11 in amastigotes is still a subject of controversy. In this
study, a strain of L. tropica was isolated, cultivated, and genotyped. The expression of
KMP-11 gene in this strain was evaluated in promastigotes and in amastigotes by RT-PCR
using specific primer pairs. The results proved the presence of mRNA of KMP-11 in both
promastigotes and amastigotes forms of L. tropica. The expression of this molecule in
amastigotes is consistent with the previously demonstrated immunoprotective capacity of
KMP-11 DNA vaccine as well as the presence of humoral and cellular immune responses
against KMP-11 in Leishmania-infected animals.
The aim of this work was determining the role of
Protein kinase C (PKC) signaling pathway in apoptosis of
Leishmania tropica promastigotes, through studying effects of PKC
specific inhibitor Staurosporine (STS) on the parasite. Our results
showed
that treated L. tropica with Staurosporine was
enough to inhibit PKC pathway completely, while viability test
showed that the concentration of STS to inhibit cell growth by 50%
(IC50 = 2.146μM).
In our study, the
presence and expression of P27 gene in L.tropica evaluated in
promastigotes by PCR and RT-PCR using specific primer pairs
that manually designed after determine the consensus sequence of
forward and reverse region of primers bet
ween species of
Leishmania by using MUSCLE (Multiple Sequence Comparison by
Log- Expectation) tool for alignment. The results proved the
presence of P27 gene in Leishmania tropica, beside presence the
mRNA of P27 gene that confirm that P27 expressed in
promastigote form of L.tropica
Leishmaniasis spreads in eighty-eight countries, including the Syrian Arab
Republic. This disease is caused by a protozoic parasite of the genus
Leishmania. Twenty-two species of leishmania were reported to be pathogenic
for human. The disease is
presented in three clinical forms: cutaneous,
mucocutaneous and visceral leishmaniasis. A set of methods for diagnosis and
identifying the type of parasite have been developed in order to give
appropriate treatment. These methods include isoenzyme analysis, serological
and immunological methods and DNA hybridization. The polymerase chain
reaction (PCR), using genomic or kinetoplastid DNA; provides an excellent tool
for diagnosis and characterization of leishmania species. In this work, the two
types of DNA were extracted in one step from cultured isolates taken from
patients. We have used kinetoplastid DNA and specific primers to determine
parasite species by PCR. These primers amplify species specific fragment. In
this study, we have determined that the causative species of the cutaneous
leishmaniasis in all patients was L.tropica.
This study was performed on blood samples of 60 people [21females, 39
males]which have visited the center of leishmaniasis in lattakia city, and their infection
with cutaneous leishmaniasis was confirmed. The people were divided into three age
gro
ups where each group contained 20 patients, the first group [10-18 years,3females and
17males], the second group [19-30 years,8 females,12males],and the third group [31-
50years,10 females and 10males]. The following tests were evaluated to each blood
sample: total white blood cell count, differential white blood cells count, hematocrit test,
and blood group type test.
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