scholarly journals Investigating the Existence of Ribosomal Protein L5 Gene in Syrian Strain of Leishmania tropica Genome: Sequencing It and Evaluating Its Immune Response as DNA Vaccine

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Mohammad Maarouf ◽  
Alyaa A. Abdlwahab
Keyword(s):  
Lymph Nodes ◽  
Ribosomal Protein ◽  
Dna Vaccine ◽  
Parasite Burden ◽  
Leishmania Tropica ◽  
Ifn Γ ◽  
Ribosomal Protein L5 ◽  
Protective Strategies ◽  
Efficacious Vaccine ◽  
Draining Lymph Nodes

Cutaneous leishmaniasis in Syria is caused mainly by Leishmania tropica. It represents a serious health problem, which has aggravated further after the civil war in the country. Until now, there are no effective protective strategies, safe therapy, or efficacious vaccine to protect from this infection. DNA vaccines represent a promising approach for achieving protection against leishmaniasis. The L5 ribosomal protein plays fundamental roles in the assembly process of the ribosome subunits, so this study has chosen the ribosomal protein L5 gene to design a DNA vaccine against Leishmania tropica infection. After proving the existence of the ribosomal protein L5 gene in a Syrian strain of Leishmania tropica (LCED Syrian 01), it was sequenced and cloned into a pCI plasmid, and the designed DNA vaccine was administered to BALB/c mice. The protective response was evaluated by measuring lesion development in immunized BALB/c mice for 6 weeks after challenging mice with the parasite. RT-qPCR was used to quantify IL-12, IFN-γ, and IL-4 in draining lymph nodes (DLNs) of immunized mice. In the final week, the parasite burden was determined in footpad lesions and local draining lymph nodes (DLNs). This study demonstrated the presence and expression of the ribosomal protein L5 gene in the Syrian strain of Leishmania tropica promastigotes. The sequence of the ribosomal protein cDNA L5 gene was determined and published in Genbank. The gene size was 918 bp. Expression was also demonstrated at the level of cDNA. This study also demonstrated that vaccination with the ribosomal protein L5 gene induces TH1 response in immunized mice. This response prevents the partial development of a skin lesion of Leishmania.

scholarly journals Intranasal Vaccination against Cutaneous Leishmaniasis with a Particulated Leishmanial Antigen or DNA Encoding LACK

2004 ◽  
Vol 72 (8) ◽  
pp. 4521-4527 ◽  
Author(s):  
Eduardo Fonseca Pinto ◽  
Roberta Olmo Pinheiro ◽  
Alice Rayol ◽  
Vicente Larraga ◽  
Bartira Rossi-Bergmann
Keyword(s):  
Lymph Nodes ◽  
Cutaneous Leishmaniasis ◽  
Oral Delivery ◽  
Parasite Burden ◽  
Mucosal Vaccine ◽  
Effective Control ◽  
Dna Encoding ◽  
Intranasal Route ◽  
Ifn Γ ◽  
Draining Lymph Nodes

ABSTRACT We have previously demonstrated that oral delivery of a disease-promoting particulated antigen of Leishmania amazonensis (LaAg) partially protects mice against cutaneous leishmaniasis. In the present work, we sought to optimize a mucosal vaccine by using the intranasal route for delivery of different antigen preparations, including (i) LaAg, (ii) soluble recombinant p36/LACK leishmanial antigen (LACK), and (iii) plasmid DNA encoding LACK (LACK DNA). BALB/c mice that received two intranasal doses of 10 μg of LaAg and were challenged 1 week postvaccination with L. amazonensis developed delayed but effective control of lesion growth. A diminished parasite burden was accompanied by enhancement of both gamma interferon (IFN-γ) and interleukin-10 levels in the lesion-draining lymph nodes. The vaccine efficacy improved with time. At 4 months postvaccination, when a strong parasite-specific TH1-type response was present in vivo, the infection was controlled for at least 5 months after challenge. In contrast to the particulated LaAg, soluble LACK (10 μg/dose) had no effect. Interestingly, LACK DNA (30 μg/dose), but not empty DNA, promoted rapid and durable protective immunity. Parasite growth was effectively controlled, and at 5 months after challenge LACK-reactive cells in both the mucosal and lesion-draining lymph nodes produced high levels of IFN-γ. These results demonstrate for the first time the feasibility of using the intranasal route for long-lived memory vaccination against cutaneous leishmaniasis with adjuvant-free crude antigens or DNA.

scholarly journals The Leishmania infantum Acidic Ribosomal Protein P0 Administered as a DNA Vaccine Confers Protective Immunity to Leishmania major Infection in BALB/c Mice

2003 ◽  
Vol 71 (11) ◽  
pp. 6562-6572 ◽  
Author(s):  
Salvador Iborra ◽  
Manuel Soto ◽  
Javier Carrión ◽  
Ana Nieto ◽  
Edgar Fernández ◽  
...  
Keyword(s):  
Ribosomal Protein ◽  
Leishmania Infantum ◽  
Leishmania Major ◽  
Parasite Burden ◽  
Immunological Response ◽  
Humoral Response ◽  
Ifn Γ ◽  
Immunogenic Properties ◽  
Ribosomal Protein P0 ◽  
Acidic Ribosomal Protein

ABSTRACT In this study, we examined the immunogenic properties of the Leishmania infantum acidic ribosomal protein P0 (LiP0) in the BALB/c mouse model. The humoral and cellular responses induced by the administration of the LiP0 antigen, either as soluble recombinant LiP0 (rLiP0) or as a plasmid DNA formulation (pcDNA3-LiP0), were determined. Also, the immunological response associated with a prime-boost strategy, consisting of immunization with pcDNA3-LiP0 followed by a boost with rLiP0, was assayed. Immunization with rLiP0 induced a predominant Th2-like humoral response, but no anti-LiP0 antibodies were induced after immunization with pcDNA3-LiP0, whereas a strong humoral response consisting of a mixed immunoglobulin G2a (IgG2a)-IgG1 isotype profile was induced in mice immunized with the prime-boost regime. For all three immunization protocols, rLiP0-stimulated production of gamma interferon (IFN-γ) in both splenocytes and lymph node cells from immunized mice was observed. However, it was only when mice were immunized with pcDNA3-LiP0 that noticeable protection against L. major infection was achieved, as determined by both lesion development and parasite burden. Immunization of mice with LiP0-DNA primes both CD4+ and CD8+ T cells, which, with the L. major challenge, were boosted to produce significant levels of IL-12-dependent, antigen-specific IFN-γ. Taken together, these data indicate that genetic vaccination with LiP0 induces protective immunological effector mechanisms, yet the immunological response elicited by LiP0 is not sufficient to keep the infection from progressing.

scholarly journals Subcutaneous immunization against Leishmania major - infection in mice: efficacy of formalin-killed promastigotes combined with adjuvants

2010 ◽  
Vol 52 (2) ◽  
pp. 95-100 ◽  
Author(s):  
Joshua M. Mutiso ◽  
John C. Macharia ◽  
Rosemary M. Mutisya ◽  
Evans Taracha
Keyword(s):  
Cutaneous Leishmaniasis ◽  
Leishmania Major ◽  
Immune Status ◽  
Parasite Burden ◽  
Interferon Γ ◽  
Cutaneous Lesions ◽  
Ifn Γ ◽  
Efficacious Vaccine ◽  
Subcutaneous Immunization ◽  
Murine Cutaneous Leishmaniasis

Formalin-killed promastigotes (FKP) of Leishmania major, in combination with Montanide ISA 720 (MISA), BCG or alum were used in vaccination of an inbred murine model against cutaneous leishmaniasis (CL). Significant and specific increases in anti-FKP IgG responses were detected for both alum-FKP and BCG-FKP compared to MISA-FKP (p < 0.001). Significant increases in splenic lymphocyte recall proliferation was obtained in the MISA-FKP vaccinated mice compared to alum-FKP or BCG-FKP vaccinated groups (p < 0.01). The highest interferon-γ responses were observed in the BCG-FKP group followed by the MISA-FKP while the alum-FKP gave the least responses. Significantly reduced lesion sizes were obtained in the MISA-FKP group compared to the BCG/alum adjuvants-FKP vaccinated groups. Although the BCG-FKP group showed the highest IFN-γ responses, it failed to control cutaneous lesions. Significant reductions in parasite numbers were observed in the MISA-FKP and BCG-FKP vaccinated groups (p < 0.001). There was a good correlation between parasite burden and IFN-γ level indicating IFN-γ response as a sensitive parameter of the immune status. In conclusion, MISA-FKP is the most efficacious vaccine formulation against murine cutaneous leishmaniasis.

Immunogenicity and Persistence of a Targeted Anti-caries DNA Vaccine

2006 ◽  
Vol 85 (10) ◽  
pp. 915-918 ◽  
Author(s):  
Q.A. Xu ◽  
F. Yu ◽  
M.W. Fan ◽  
Z. Bian ◽  
Z. Chen ◽  
...  
Keyword(s):  
Immune Responses ◽  
Lymph Nodes ◽  
Dna Vaccine ◽  
Antibody Responses ◽  
Cervical Lymph Nodes ◽  
Booster Immunization ◽  
Inoculation Site ◽  
The Us ◽  
Kinetics Of ◽  
Draining Lymph Nodes

We have previously reported that a targeted anti-caries DNA vaccine, pGJA-P, induced accelerated and increased antibody responses compared with a non-targeted anti-caries DNA vaccine. Recently, pGJA-P/VAX, a new targeted anti-caries DNA vaccine for human trials, was constructed by replacing the pCI vector used in the construction of pGJA-P with pVAX1, the only vector authorized by the US Food and Drug Administration in clinical trials. Here, we report on our exploration of the kinetics of the antibody responses generated following pGJA-P/VAX immunization and the persistence of pGJA-P/VAX at both the inoculation site and the draining lymph nodes. Intranasal vaccination of mice with pGJA-P/VAX induced strong antibody responses that lasted for more than 6 months. Furthermore, pGJA-P/VAX could still be detected at both the inoculation site and the draining cervical lymph nodes 6 months after immunization. Thus, the persistent immune responses are likely due to the DNA depot in the host, which acts as a booster immunization.

Determination of a CD4+CD25−FoxP3+ T cells subset in tumor-draining lymph nodes of colorectal cancer secreting IL-2 and IFN-γ

Tumor Biology ◽  
2016 ◽  
Vol 37 (11) ◽  
pp. 14659-14666 ◽  
Author(s):  
Morteza Jafarinia ◽  
Fereshteh Mehdipour ◽  
Seyed Vahid Hosseini ◽  
Leila Ghahramani ◽  
Masood Hosseinzadeh ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
T Cells ◽  
Lymph Nodes ◽  
Ifn Γ ◽  
Draining Lymph Nodes

scholarly journals Enrichment of Innate Lymphoid Cell Populations in Gingival Tissue

2018 ◽  
Vol 97 (12) ◽  
pp. 1399-1405 ◽  
Author(s):  
J.L. Brown ◽  
L. Campbell ◽  
J. Malcolm ◽  
A. Adrados Planell ◽  
J.P. Butcher ◽  
...  
Keyword(s):  
Oral Cavity ◽  
Lymph Nodes ◽  
Population Group ◽  
Lymphoid Cells ◽  
Gingival Tissue ◽  
Regional Lymph Nodes ◽  
Mucosal Surfaces ◽  
Ifn Γ ◽  
Draining Lymph Nodes ◽  
Group 1

Innate lymphoid cells (ILCs) are a population of lymphocytes that act as the first line of immunologic defense at mucosal surfaces. The ILC family in the skin, lungs, and gastrointestinal tissues has been investigated, and there are reports of individual subsets of ILCs in the oral tissues. We sought to investigate the whole ILC population (group 1, 2, and 3 subsets) in the murine gingivae and the lymph nodes draining the oral cavity. We show that ILCs made up a greater proportion of the whole CD45+ lymphocyte population in the murine gingivae (0.356% ± 0.039%) as compared with the proportion of ILCs in the draining lymph nodes (0.158% ± 0.005%). Cytokine profiling of the ILC populations demonstrated different proportions of ILC subsets in the murine gingivae versus the regional lymph nodes. The majority of ILCs in the draining lymph nodes expressed IL-5, whereas there were equal proportions of IFN-γ- and IL-5 expressing ILCs in the oral mucosa. The percentage of IL-17+ ILCs was comparable between the murine gingivae and the oral draining lymph nodes. These data suggest an enrichment of ILCs in the murine gingivae, and these ILCs reflect a cytokine profile discrepant to that of the local draining lymph nodes. These studies indicate diversity and enrichment of ILCs at the oral mucosal surface. The function of ILCs in the oral cavity remains to be determined; here, we provide a premise of ILC populations that merits future consideration in investigations of mouse models and human tissues.

scholarly journals Gene Transcription Profile in Mice Vaccinated with Ultraviolet-attenuated Cercariae of Schistosoma japonicum Reveals Molecules Contributing to Elevated IFN-γ Levels

2005 ◽  
Vol 37 (4) ◽  
pp. 254-264 ◽  
Author(s):  
Xiang Zhu ◽  
Zhao-Song Zhang ◽  
Min-Jun Ji ◽  
Hai-Wei Wu ◽  
Yong Wang ◽  
...  
Keyword(s):  
Lymph Nodes ◽  
Schistosoma Japonicum ◽  
Protective Immunity ◽  
Early Stage ◽  
Post Vaccination ◽  
Ifn Γ ◽  
Preliminary Study ◽  
Gene Transcription Profile ◽  
Th1 And Th2 Responses ◽  
Draining Lymph Nodes

AbstractVaccination with ultraviolet-attenuated cercariae of Schistosoma japonicum induced protective immunity against challenge infection in experimental animal models. Our preliminary study on the transcription levels of IFN-γ and IL-4 in splenic CD4+ T cells revealed that attenuated cercariae elicited predominantly a Th1 response in mice at the early stage, whereas normal cercariae stimulated primarily Independent responses. Further analysis on the gene profile of the skin-draining lymph nodes demonstrated that the levels of IFN-γ were significantly higher in vaccinated mice than those in infected mice at day 4, 7 and 14 post-vaccination or post-infection. However, for IL-12 and IL-4, the potent inducers of Th1 and Th2 responses, respectively, as well as IL-10, there were no differences over the course of the experiment between the infected and vaccinated mice. To explore the underlying factors that may potentially contribute to elevated IFN-γ in vaccinated mice, the mRNA profiles of the skin-draining lymph nodes at day 4 post-exposure were compared using oligonucleotide microarrays. Within the 847 probe sets with increased signal values, we focused on chemokines, cytokines and relevant receptors, which were validated by semi-quantitative RT-PCR. A comprehensive understanding of the immune mechanisms of attenuated cercariae-induced protection may contribute to developing efficient vaccination strategies against S. japonicum, especially during the early stage of infection.

scholarly journals Immunogenicity and potential protection of DNA vaccine of Leishmania martiniquensis against Leishmania infection in mice

2021 ◽  
Vol 15 (09) ◽  
pp. 1328-1338
Author(s):  
Thuntawat Aunguldee ◽  
Orapin Gerdprasert ◽  
Piyatida Tangteerawatana ◽  
Amporn Jariyapongskul ◽  
Saovanee Leelayoova ◽  
...  
Keyword(s):  
Dna Vaccine ◽  
Consensus Sequence ◽  
Parasite Burden ◽  
Interleukin 10 ◽  
Leishmania Infection ◽  
Control Group ◽  
Ifn Γ ◽  
Polymerase Chain ◽  
Immunocompromised Hosts ◽  
Partial Consensus

Introduction: In Thailand, Leishmania martiniquensis is the predominant species causing cutaneous and visceral leishmaniasis. Its incidence has been increasing among immunocompetent and immunocompromised hosts. We developed a prototype DNA vaccine using a partial consensus sequence of the cysteine protease B (cpb) gene derived from L. martiniquensis from Thai patients. Methodology: The laboratory inbred strain of albino BALB/c mice were immunized intramuscularly three times at 2-week intervals (weeks 0, 2, and 4) with cpb plasmid DNA (pcDNA_cpb) with or without the adjuvant, monoolein (pcDNA_cpb-MO). Mice were challenged at week 6 with L. martiniquensis promastigotes. Sera were analysed for IgG1, IgG2a, interferon gamma and interleukin 10 (IFN-γ and IL-10, respectively) levels at weeks 0, 4, and 9. Additionally, livers and spleens were also analysed for parasite burden using immunohistochemistry and real-time polymerase chain (qPCR) assays. Results: Three weeks after promastigote challenge, vaccinated mice showed significantly increased levels of IgG2a and IFN-γ while IL-10 level was significantly reduced when compared with those in the control group (p < 0.01). Parasite burden in the livers and spleens of vaccinated mice significantly decreased. In addition, a significant increase in mature granuloma formation in the livers when compared with those of the control group (p < 0.05) was found, indicating increased T-helper cells (Th1)-induced inflammation and destruction of amastigotes. Monoolein produced a booster effect to enhance the mouse Th1 protective immunity. Conclusions: The prototype DNA vaccine could induce a Th1 immune response that conferred potential protection to the L. martiniquensis promastigote challenge in BALB/c mice.

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