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

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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Vaccination with a DNA Vaccine Coding for Perforin-Like Protein 1 and MIC6 Induces Significant Protective Immunity against Toxoplasma gondii

Clinical and Vaccine Immunology ◽

10.1128/cvi.05578-11 ◽

2012 ◽

Vol 19 (5) ◽

pp. 684-689 ◽

Cited By ~ 26

Author(s):

Hai-Kuo Yan ◽

Zi-Guo Yuan ◽

Hui-Qun Song ◽

Eskild Petersen ◽

Yang Zhou ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Dna Vaccine ◽

Interleukin 2 ◽

Parasite Load ◽

Control Group ◽

Igg Antibody ◽

Content Type ◽

Humoral Immune ◽

Humoral Immune Responses ◽

Ifn Γ

ABSTRACTHost cell invasion byToxoplasma gondiiis tightly related to microneme protein 6 (MIC6) andT. gondiiperforin-like protein 1 (TgPLP1). In this study, we constructed a DNA vaccine expressing a TgPLP1/MIC6 fusion protein using the pIRESneo vector, and we evaluated the immune response induced by this vaccine in Kunming mice. Levels of IgG antibody, gamma interferon (IFN-γ), interleukin 2 (IL-2), IL-12, IL-4, and IL-10 were examined. Five mice were chosen randomly from every group (vaccinated groups or the nonvaccinated control group) and were challenged intragastrically with 80 cysts ofT. gondiistrain PRU (genotype II) in order to observe mortality daily. To analyze protection against a less-virulent challenge, eight mice of each group were orally infected with 20 cysts of strain PRU at the 14th day after the last immunization. The brain parasite load was evaluated 6 weeks after infection. The results demonstrated that immunization with pIRESneo/MIC6/PLP1 resulted in the lowest brain cyst count and prolonged the survival time of immunized mice. The levels ofToxoplasma-specific IgG, IFN-γ, IL-2, and IL-12 increased significantly, and the numbers of cysts in brains decreased more obviously, in the group immunized with plasmid pIRESneo/MIC6/PLP1 than in the other groups (P< 0.05). Compared with pIRESneo/MIC6/PLP1, coimmunization with pIRESneo/MIC6/PLP1 and adjuvant murine IL-18 promoted cellular and humoral immune responses but did not contribute significantly to cyst reduction (65.43% versus 61.60%) or the survival of immunized mice (45.0 ± 2.9 days versus 42.8 ± 2.9 days) (P> 0.05). Furthermore, the study also showed that the immune efficacy induced by pIRESneo/MIC6/PLP1 was better than that induced by pVAX/PLP1 or pVAX/MIC6 alone.

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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

Journal of Parasitology Research ◽

10.1155/2021/6617270 ◽

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.

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Ex Vivo Generation of Donor Antigen-Specific Immunomodulatory Cells

Cell Transplantation ◽

10.1177/0963689718794642 ◽

2018 ◽

Vol 27 (11) ◽

pp. 1692-1704

Author(s):

M. Watanabe ◽

Makiko Kumagai-Braesch ◽

M. Yao ◽

S. Thunberg ◽

D. Berglund ◽

...

Keyword(s):

T Cells ◽

Mononuclear Cells ◽

Ex Vivo ◽

Human Peripheral Blood ◽

Human Leukocyte ◽

Interleukin 10 ◽

Control Group ◽

Peripheral Blood Mononuclear ◽

Foxp3 Mrna ◽

Ifn Γ

Adoptive transfer of alloantigen-specific immunomodulatory cells generated ex vivo with anti-CD80/CD86 mAbs (2D10.4/IT2.2) holds promise for operational tolerance after transplantation. However, good manufacturing practice is required to allow widespread clinical application. Belatacept, a clinically approved cytotoxic T-lymphocyte antigen 4-immunoglobulin that also binds CD80/CD86, could be an alternative agent for 2D10.4/IT2.2. With the goal of generating an optimal cell treatment with clinically approved reagents, we evaluated the donor-specific immunomodulatory effects of belatacept- and 2D10.4/IT2.2-generated immunomodulatory cells. Immunomodulatory cells were generated by coculturing responder human peripheral blood mononuclear cells (PBMCs) (50 × 106 cells) with irradiated donor PBMCs (20 × 106 cells) from eight human leukocyte antigen-mismatched responder–donor pairs in the presence of either 2D10.4/IT2.2 (3 μg/106 cells) or belatacept (40 μg/106 cells). After 14 days of coculture, the frequencies of CD4+ T cells, CD8+ T cells, and natural killer cells as well as interferon gamma (IFN-γ) production in the 2D10.4/IT2.2- and belatacept-treated groups were lower than those in the control group. The percentage of CD19+ B cells was higher in the 2D10.4/IT2.2- and belatacept-treated groups than in the control group. The frequency of CD4+CD25+CD127lowFOXP3+ T cells increased from 4.1±1.0% (preculture) to 7.1±2.6% and 7.3±2.6% (day 14) in the 2D10.4/IT2.2- and belatacept-treated groups, respectively ( p<0.05). Concurrently, delta-2 FOXP3 mRNA expression increased significantly. Compared with cells derived from the no-antibody treated control group, cells generated from both the 2D10.4/IT2.2- and belatacept-treated groups produced lower IFN-γ and higher interleukin-10 levels in response to donor-antigens, as detected by enzyme-linked immunospot. Most importantly, 2D10.4/IT2.2- and belatacept-generated cells effectively impeded the proliferative responses of freshly isolated responder PBMCs against donor-antigens. Our results indicate that belatacept-generated donor-specific immunomodulatory cells possess comparable phenotypes and immunomodulatory efficacies to those generated with 2D10.4/IT2.2. We suggest that belatacept could be used for ex vivo generation of clinical grade alloantigen-specific immunomodulatory cells for tolerance induction after transplantation.

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Immunogenicity and Protective Efficacy against Murine Tuberculosis of a Prime-Boost Regimen with BCG and a DNA Vaccine Expressing ESAT-6 and Ag85A Fusion Protein

Clinical and Developmental Immunology ◽

10.1155/2011/617892 ◽

2011 ◽

Vol 2011 ◽

pp. 1-10 ◽

Cited By ~ 22

Author(s):

Jia Lu ◽

Chun Wang ◽

Zhiguang Zhou ◽

Ying Zhang ◽

Tingting Cao ◽

...

Keyword(s):

Fusion Protein ◽

Dna Vaccine ◽

Protective Efficacy ◽

Bacterial Load ◽

Control Group ◽

Cell Mediated Immunity ◽

Significant Protection ◽

Booster Vaccine ◽

Ifn Γ ◽

Negative Population

Heterologous prime-boost regimens utilizing BCG as a prime vaccine probably represent the best hope for the development of novel tuberculosis (TB) vaccines. In this study, we examined the immunogenicity and protective efficacy of DNA vaccine (pcD685A) expressing the fusion protein of Ag85A and ESAT-6 (r685A) and its booster effects in BCG-immunized mice. The recombinant r685A fusion protein stimulated higher level of antigen-specific IFN-γ release in tuberculin skin test- (TST-) positive healthy household contacts of active pulmonary TB patients than that in TST-negative population. Vaccination of C57BL/6 mice with pcD685A resulted in significant protection against challenge with virulentMycobacterium tuberculosisH37Rv when compared with the control group. Most importantly, pcD685A could act as a BCG booster and amplify Th1-type cell-mediated immunity in the lung of BCG-vaccinated mice as shown the increased expression of IFN-γ. The most significant reduction in bacterial load of both spleen and lung was obtained in mice vaccinated with BCG prime and pcD685A DNA booster when compared with BCG or pcD685A alone. Thus, our study indicates that pcD685A may be an efficient booster vaccine against TB with a strong ability to enhance prior BCG immunity.

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THE EFFECT OF ALPHA-MANGOSTIN IN BALANCING THE RATIO OF CYTOKINES PRO-AND ANTI-INFLAMMATION-GAMMA (IFN-γ/IL-10) AND SEVERITY OF THE DISEASE IN MICE INFECTED WITH MYCOBACTERIUM TUBERCULOSIS MULTIDRUG-RESISTANT

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2016.v9s3.14544 ◽

2016 ◽

Vol 9 (9) ◽

pp. 273

Author(s):

Dyan Kunthi Nugrahaeni ◽

Suharyo Hadisaputro ◽

Ari Suwondo ◽

Edi Dharmana

Keyword(s):

Mycobacterium Tuberculosis ◽

Interleukin 10 ◽

Multidrug Resistant ◽

Interferon Γ ◽

Control Group ◽

Cytokine Levels ◽

Ifn Γ ◽

Severity Of The Disease ◽

And Control ◽

Anti Inflammation

ABSTRACTObjective: The objective of this study was observed to measure the effect of alpha-mangostin in balancing the ratio of interferon-gamma (IFN-γ) andinterleukin-10 (IL-10), and the severity of the disease in mice which infected with Mycobacterium tuberculosis multidrug-resistant (TB-MDR).Method: Infected BALB/c mice were consisted of five groups: Treated with anti-TB drugs+α-mangostin, treated with anti-TB drugs, given α-mangostinduring treatment, and control group. Cytokine levels of culture supernatant of spleen cells were measured by enzyme-linked immunosorbent assay.The number of bacterial colonies was derived from a primary cell culture of bronchoalveolar lavage. Statistical analysis was performed with Anova,Kruskal-Wallis test and correlation Pearson, and Spearman-rank test.Result: Median IFN-γ production was higher in mice, which given with α-mangostin during treatment is 1838.2 pg/ml and control is 1585.5 pg/mlcompared treated with anti-TB drugs+α-mangostin (1312 pg/ml) and anti-TB drugs (1429.3 pg/ml) (p>0.05). The highest result production of medianIL-10 in the 3th group is (465.91 pg/ml) and the lowest in the control group is 195.29 pg/ml, p>0.05. Median IFN-γ/IL-10 ratio of the 3th group verylow (3.94), it means the 3th group is experienced with severity of TB. Alpha-mangostin was decreased in severity of disease based on the number ofTB-MDR bacterial colonies (p≤0.05).Conclusion: α-mangostin have an effect on the balancing IFN-γ/IL-10 ratio and reduce a severity of TB-MDR with using immunomodulator.Keywords: Tuberculosis multidrug-resistant, Alpha-mangostin, Interferon-γ/interleukin-10 ratio, Number of tuberculosis multidrug-resistantbacterial colonies.

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Peptide-based vaccine successfully induces protective immunity against canine visceral leishmaniasis

npj Vaccines ◽

10.1038/s41541-019-0144-2 ◽

2019 ◽

Vol 4 (1) ◽

Cited By ~ 4

Author(s):

Elodie Petitdidier ◽

Julie Pagniez ◽

Joana Pissarra ◽

Philippe Holzmuller ◽

Gérard Papierok ◽

...

Keyword(s):

Visceral Leishmaniasis ◽

Protective Immunity ◽

Vaccine Candidate ◽

Parasite Burden ◽

Control Group ◽

Preclinical Trial ◽

Ifn Γ ◽

Secretory Products ◽

Carboxy Terminal ◽

Zoonotic Visceral Leishmaniasis

AbstractDogs are the main reservoir of zoonotic visceral leishmaniasis. Vaccination is a promising approach to help control leishmaniasis and to interrupt transmission of the Leishmania parasite. The promastigote surface antigen (PSA) is a highly immunogenic component of Leishmania excretory/secretory products. A vaccine based on three peptides derived from the carboxy-terminal part of Leishmania amazonensis PSA and conserved among Leishmania species, formulated with QA-21 as adjuvant, was tested on naive Beagle dogs in a preclinical trial. Four months after the full course of vaccination, dogs were experimentally infected with Leishmania infantum promastigotes. Immunization of dogs with peptide-based vaccine conferred immunity against experimental infection with L. infantum. Evidence for macrophage nitric oxide production and anti-leishmanial activity associated with IFN-γ production by lymphocytes was only found in the vaccinated group. An increase in specific IgG2 antibodies was also measured in vaccinated dogs from 2 months after immunization. Additionally, after challenge with L. infantum, the parasite burden was significantly lower in vaccinated dogs than in the control group. These data strongly suggest that this peptide-based vaccine candidate generated cross-protection against zoonotic leishmaniasis by inducing a Th1-type immune response associated with production of specific IgG2 antibodies. This preclinical trial including a peptide-based vaccine against leishmaniasis clearly demonstrates effective protection in a natural host. This approach deserves further investigation to enhance the immunogenicity of the peptides and to consider the possible engineering of a vaccine targeting several Leishmania species.

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DNA Vaccine Using Mycobacterium bovis Ag85B Antigen Induces Partial Protection against Experimental Infection in BALB/c Mice

Clinical and Vaccine Immunology ◽

10.1128/cvi.00151-06 ◽

2006 ◽

Vol 13 (8) ◽

pp. 930-935 ◽

Cited By ~ 15

Author(s):

Francisco M. Teixeira ◽

Henrique C. Teixeira ◽

Ana Paula Ferreira ◽

Michele F. Rodrigues ◽

Vasco Azevedo ◽

...

Keyword(s):

Dna Vaccine ◽

Mycobacterium Bovis ◽

Vaccine Development ◽

Virulent Strain ◽

Intracellular Cytokine Staining ◽

Bacterial Load ◽

Interleukin 4 ◽

Control Group ◽

Partial Protection ◽

Ifn Γ

ABSTRACT Bovine tuberculosis is a major cause of economic loss in countries where it is endemic, and in some countries, it may be a significant zoonotic disease problem. Therefore, new strategies for vaccine development are required, and among them, genetic immunization has potential value. The main goal of this study was to test the Mycobacterium bovis Ag85B gene as a DNA vaccine following challenge with an M. bovis virulent strain (ATCC 19274). Groups of BALB/c mice (n = 10) were immunized four times intramuscularly with the pCI-Ag85B construct or the pCI vector alone as the control. High titers of total immunoglobulin G (IgG), IgG1, and IgG2a anti-Ag85B were measured in pCI-Ag85B immunized mice when compared to the pCI control group. Regarding cellular immunity, significant levels of gamma interferon (IFN-γ) (1,100 ± 157 pg/ml) and tumor necrosis factor alpha (650 ± 42 pg/ml) but not interleukin-4 were detected in splenocyte culture supernatants of pCI-Ag85B-vaccinated mice following stimulation with recombinant Ag85B. Further, the main source of IFN-γ is CD8+ T cells, as demonstrated by intracellular cytokine staining. As far as protection, a significant reduction in bacterial load in spleens (P < 0.05) was detected in pCI-Ag85B-immunized mice compared to the pCI vector control group. The results obtained here suggest that use of the Ag85B DNA vaccine is a promising strategy to control M. bovis infection due to its ability to induce a Th1 type of immune response. However, protective efficacy needs to be improved, since partial protection was achieved in spleens but not in lungs of vaccinated mice.

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Immunization with the HisAK70 DNA Vaccine Induces Resistance against Leishmania Amazonensis Infection in BALB/c Mice

Vaccines ◽

10.3390/vaccines7040183 ◽

2019 ◽

Vol 7 (4) ◽

pp. 183 ◽

Cited By ~ 1

Author(s):

Abel Martínez-Rodrigo ◽

Daniel S. Dias ◽

Patrícia A. F. Ribeiro ◽

Bruno M. Roatt ◽

Alicia Mas ◽

...

Keyword(s):

Dna Vaccine ◽

Leishmania Major ◽

Parasite Species ◽

Leishmania Amazonensis ◽

Control Group ◽

Gm Csf ◽

Ifn Γ ◽

Cellular Immune ◽

Cutaneous Leishmaniosis ◽

Type Response

Leishmania amazonensis is the aetiological agent of a broad spectrum of leishmaniosis in South America. It can cause not only numerous cases of cutaneous leishmaniosis but also diffuse cutaneous leishmaniosis. Considering the diversity of parasite species causing different forms of the disease that coexist in the same region, it is desirable to develop a vaccine capable of eliciting cross-protection. We have previously described the use of HisAK70 DNA vaccine for immunization of mice to assess the induction of a resistant phenotype against Leishmania major and infantum infections. In this study, we extended its application in the murine model of infection by using L. amazonensis promastigotes. Our data revealed that 14 weeks post-infection, HisAK70-vaccinated mice showed key biomarkers of protection, such as higher iNOS/arginase activity, IFN-γ/IL-10, IFN-γ/IL-4, and GM-CSF/IL-10 ratios, in addition to an IgG2a-type response when compared to the control group. These findings correlated with the presentation of lower footpad swelling and parasite burdens in the immunized compared to the control mice. Overall, this study suggests that immunization with HisAK70 may be considered a suitable tool to combat leishmaniosis as it is able to induce a potent cellular immune response, which allows to control the infection caused by L. amazonensis.

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A Mutant of Staphylococcal Enterotoxin C Devoid of Bacterial Superantigenic Activity Elicits a Th2 Immune Response for Protection against Staphylococcus aureus Infection

Infection and Immunity ◽

10.1128/iai.73.1.174-180.2005 ◽

2005 ◽

Vol 73 (1) ◽

pp. 174-180 ◽

Cited By ~ 17

Author(s):

Dong-Liang Hu ◽

Jing-Chun Cui ◽

Katsuhiko Omoe ◽

Hiroshi Sashinami ◽

Yuichi Yokomizo ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Neutralizing Antibodies ◽

Interleukin 10 ◽

Staphylococcal Enterotoxin ◽

Control Group ◽

Necrosis Factor Alpha ◽

T Helper 2 ◽

Th2 Immune Response ◽

Ifn Γ

ABSTRACT Staphylococcal enterotoxin C (SEC), a bacterial superantigenic exotoxin, is commonly produced by invasive Staphylococcus aureus isolates, especially methicillin-resistant strains and isolates from animal diseases. We constructed and expressed a nontoxic mutant SEC (mSEC) and investigated whether immunization with mSEC, which is devoid of superantigenic activity, can protect against S. aureus infection. Mice were immunized with mSEC and challenged with viable S. aureus. The bacterial counts in the organs of mSEC-immunized mice were significantly lower and the survival rate was higher than the corresponding values for the control group. Immunization with mSEC strongly induced the production of T-helper 2 type antibodies, immunoglobulin G1, and immunoglobulin G2b. The production of interleukin-10 (IL-10) and IL-4 was significantly greater in immunized mice challenged with S. aureus than in the control mice, whereas the production of gamma interferon (IFN-γ) was significantly decreased in the immunized mice. The cytokine response in a spleen cell culture that was stimulated with heat-killed S. aureus or SEC showed that immunization with mSEC inhibited IFN-γ production and up-regulated IL-10 production in vitro. Furthermore, IFN-γ and tumor necrosis factor alpha production in vitro was significantly inhibited by sera from mSEC-immunized mice but not by sera from control mice. These results suggest that immunization with mSEC devoid of superantigenic properties provides protection against S. aureus infection and that the protection might be mediated by SEC-specific neutralizing antibodies.

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Protection of Mice with a Divalent Tuberculosis DNA Vaccine Encoding Antigens Ag85B and MPT64

Acta Biochimica et Biophysica Sinica ◽

10.1093/abbs/36.4.269 ◽

2004 ◽

Vol 36 (4) ◽

pp. 269-276 ◽

Cited By ~ 13

Author(s):

Xia Tian ◽

Hong Cai ◽

Yu-Xian Zhu

Keyword(s):

Dna Vaccine ◽

Protective Efficacy ◽

Vaccine Group ◽

Control Group ◽

Lung Weight ◽

Promising Tool ◽

Ifn Γ ◽

Immunodominant Antigens ◽

High Doses ◽

Tuberculosis Disease

Abstract DNA vaccine may be a promising tool for controlling tuberculosis development. However, vaccines encoding single antigens of mycobacterium did not produce protective effect as BCG did. In the present study, we evaluated the immunogenicity and protective efficacy of a divalent DNA vaccine encoding two immunodominant antigens Ag85B and MPT64 of Mycobacterium tuberculosis. We found that both humoral and Th1-type (high IFN-γ, low IL-4) cellular responses obtained from the divalent DNA vaccine group were significantly higher than that conferred by BCG. RT-PCR results showed that antigens were expressed differentially in various organs in divalent DNA vaccine group. The survival rate for mice treated with the divalent DNA vaccine after challenging with high doses of virulent M. tuberculosis H37Rv was significantly higher than that of the BCG group or any of the single DNA vaccine group. Significant differences were also found between the single and divalent DNA vaccinated mice in terms of body, spleen and lung weight. Bacterial loading decreased about 2000-fold in lungs and about 100-fold in spleens of divalent DNA vaccinated mice when compared with that of the control group. We conclude that our divalent DNA vaccine may be a better choice for controlling tuberculosis disease in animals.

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