Eimeria maxima Rhomboid-like Protein 5 Provided Partial Protection against Homologous Challenge in Forms of Recombinant Protein and DNA Plasmid in Chickens

Eimeria maxima (E. maxima) is one of the most prevalent species that causes chicken coccidiosis on chicken farms. During apicomplexan protozoa invasion, rhomboid-like proteins (ROMs) cleave microneme proteins (MICs), allowing the parasites to fully enter the host cells, which suggests that ROMs have the potential to be candidate antigens for the development of subunit or DNA vaccines against coccidiosis. In this study, a recombinant protein of E. maxima ROM5 (rEmROM5) was expressed and purified and was used as a subunit vaccine. The eukaryotic expression plasmid of pVAX–EmROM5 was constructed and was used as a DNA vaccine. Chickens who were two weeks old were vaccinated with the rEmROM5 and pVAX–EmROM5 vaccines twice, with a one-week interval separating the vaccination periods. The transcription and expression of pVAX–EmROM5 in the injected sites were detected through reverse transcription PCR (RT-PCR) and Western blot (WB) assays. The cellular and humoral immune responses that were induced by EmROM5 were determined by detecting the proportion of CD4+ and CD8+ T lymphocytes, the cytokine levels, and the serum antibody levels. Finally, vaccination-challenge trials were conducted to evaluate the protective efficacy of EmROM5 in forms of the recombinant protein (rEmROM5) and in the DNA plasmid (pVAX–EmROM5) separately. The results showed that rEmROM5 was about 53.64 kDa, which was well purified and recognized by the His-Tag Mouse Monoclonal antibody and the chicken serum against E. maxima separately. After vaccination, pVAX–EmROM5 was successfully transcribed and expressed in the injected sites of the chickens. Vaccination with rEmROM5 or pVAX–EmROM5 significantly promoted the proportion of CD4+/CD3+ and CD8+/CD3+ T lymphocytes, the mRNA levels of the cytokines IFN-γ, IL-2, IL-4, IL-17, TNF SF15, and IL-10, and specific IgG antibody levels compared to the control groups. The immunization also significantly reduced the weight loss, oocyst production, and intestinal lesions that are caused by E. maxima infection. The anticoccidial index (ACI)s of the vaccinated groups were beyond 160, showing moderate protection against E. maxima infection. In summary, EmROM5 was able to induce a robust immune response and effective protection against E. maxima in chickens in the form of both a recombinant protein and DNA plasmid. Hence, EmROM5 could be used as a candidate antigen for DNA vaccines and subunit vaccines against avian coccidiosis.

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DNA vaccines expressing pneumococcal surface protein A (PspA) elicit protection levels comparable to recombinant protein

Journal of Medical Microbiology ◽

10.1099/jmm.0.46217-0 ◽

2006 ◽

Vol 55 (4) ◽

pp. 375-378 ◽

Cited By ~ 14

Author(s):

Daniela M. Ferreira ◽

Eliane N. Miyaji ◽

Maria Leonor S. Oliveira ◽

Michelle Darrieux ◽

Ana Paula M. Arêas ◽

...

Keyword(s):

Streptococcus Pneumoniae ◽

Recombinant Protein ◽

Dna Vaccines ◽

Protein A ◽

Surface Protein ◽

Humoral Response ◽

Cost Effective ◽

Promising Candidate ◽

Pneumococcal Surface Protein A ◽

Antibody Levels

Pneumococcal surface protein A (PspA) is a promising candidate for the development of cost-effective vaccines against Streptococcus pneumoniae. In the present study, BALB/c mice were immunized with DNA vaccine vectors expressing the N-terminal region of PspA. Animals immunized with a vector expressing secreted PspA developed higher levels of antibody than mice immunized with the vector expressing the antigen in the cytosol. However, both immunogens elicited similar levels of protection against intraperitoneal challenge. Furthermore, immunization with exactly the same fragment in the form of a recombinant protein, with aluminium hydroxide as an adjuvant, elicited even higher antibody levels, but this increased humoral response did not correlate with enhanced protection. These results show that DNA vaccines expressing PspA are able to elicit protection levels comparable to recombinant protein, even though total anti-PspA IgG response is considerably lower.

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Vaccination of Mice with Gonococcal TbpB Expressed In Vivo from Venezuelan Equine Encephalitis Viral Replicon Particles

Infection and Immunity ◽

10.1128/iai.74.3.1612-1620.2006 ◽

2006 ◽

Vol 74 (3) ◽

pp. 1612-1620 ◽

Cited By ~ 13

Author(s):

Christopher E. Thomas ◽

Weiyan Zhu ◽

Cornelius N. Van Dam ◽

Nancy L. Davis ◽

Robert E. Johnston ◽

...

Keyword(s):

Recombinant Protein ◽

Specific Antibody ◽

Serum Antibody ◽

Venezuelan Equine Encephalitis ◽

Equine Encephalitis Virus ◽

Secretion Signal ◽

Replicon Particle ◽

Intact Surface ◽

Antibody Levels

ABSTRACT We investigated the immunogenicity of gonococcal transferrin binding protein B (TbpB) expressed with and without a eukaryotic secretion signal from a nonpropagating Venezuelan equine encephalitis virus replicon particle (VRP) delivery system. TbpB was successfully expressed in baby hamster kidney (BHK) cells, and the presence of the eukaryotic secretion signal not only apparently increased the protein's expression but also allowed for extracellular localization and glycosylation. Mice immunized with VRPs produced significant amounts of serum antibody although less than the amounts produced by mice immunized with recombinant protein. The response of mice immunized with VRPs encoding TbpB was consistently more Th1 biased than the response of mice immunized with recombinant protein alone. Boosting with recombinant protein following immunization with TbpB VRPs resulted in higher specific-antibody levels without altering the Th1/Th2 bias. Most of the immunization groups produced significant specific antibody binding to the intact surface of the homologous Neisseria gonorrhoeae strain. Immunization with TbpB VRPs without a eukaryotic secretion signal generated no measurable specific antibodies on the genital mucosal surface, but inclusion of a eukaryotic secretion signal or boosting with recombinant protein resulted in specific immunoglobulin G (IgG) and IgA in mucosal secretions after TbpB VRP immunization. The TbpB VRP system has potential for an N. gonorrhoeae vaccine.

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Lymphocyte subpopulations and antibody levels in immunized malnourished children

British Journal Of Nutrition ◽

10.1079/bjn19820082 ◽

1982 ◽

Vol 48 (1) ◽

pp. 7-14 ◽

Cited By ~ 18

Author(s):

L. S. Salimonu ◽

A. O. K. Johnson ◽

A. I. O. Williams ◽

G. Iyabo Adeleye ◽

B. O. Osunkoya

Keyword(s):

T Lymphocytes ◽

T Lymphocyte ◽

Tetanus Toxoid ◽

Measles Virus ◽

Serum Antibody ◽

Lymphocyte Subpopulations ◽

Malnourished Children ◽

Antibody Levels ◽

And Control

1. The proportions of lymphocyte subpopulations (by rosette tests) and the serum antibody levels (using haemagglutination techniques) were estimated in malnourished and well fed Nigerian children before and up to 21 d after immunization with tetanus toxoid or measles virus vaccine.2. Significantly diminished (P < 0.01) mean percentage T lymphocyte levels and considerably higher mean percentage null cell levels were observed in the malnourished children before immunization with either of the vaccines.3. There was comparable in vivo increases in percentage T lymphocytes in malnourished and control children following the administration of each antigen.4. The mean percentage B lymphocyte levels were similar in the control and malnourished children before and after the immunization.5. There was a slight depression in the tetanus antibody levels (P > 0.2) but a significant diminution (P < 0.01) in measles virus antibody concentrations in the malnourished children.6. Rise in mean percentage T lymphocytes corresponded with the elevation in mean tetanus antibody levels in both malnourished and control children following tetanus toxoid immunization. This was however not the situation in the malnourished children following immunization with measles virus.7. The observed depressed T lymphocyte number in malnourished children may in practice affect their handling of antigens such as measles virus in vivo.

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Enhanced Murine Antigen-Specific Gamma Interferon and Immunoglobulin G2a Responses by Using Mycobacterial ESAT-6 Sequences in DNA Vaccines

Infection and Immunity ◽

10.1128/iai.71.4.2239-2243.2003 ◽

2003 ◽

Vol 71 (4) ◽

pp. 2239-2243 ◽

Cited By ~ 10

Author(s):

F. Chris Minion ◽

Sreekumar A. Menon ◽

Gregory G. Mahairas ◽

M. J. Wannemuehler

Keyword(s):

Immune Responses ◽

Dna Vaccines ◽

Serum Antibody ◽

Interleukin 10 ◽

Gamma Interferon ◽

Viral Pathogens ◽

Ifn Γ ◽

Antibody Levels

ABSTRACT The Mycobacterium tuberculosis protein ESAT-6 has unusual immune stimulating activities, has been implicated in the recall of long-lived immunity, and induces protection against tuberculosis in mice. For many diseases caused by bacterial or viral pathogens, a strong cell-mediated immune (i.e., type 1) response is often required for recovery or protection. Therefore, it is important to design immunization regimens that induce agent-specific type 1 immunity. We have shown in previous studies that ESAT-6 could enhance antigen-specific type 1 immune responses in BALB/c mice against a second antigen when presented as a purified fusion protein. It was also of interest to determine if ESAT-6 could enhance the type 1 response against a second antigen beyond that afforded by DNA vaccination through CpG motifs. This was tested by using gene fusions of ESAT-6 and the Mycoplasma hyopneumoniae surface antigen P71. Modified P71 gene sequences were cloned with or without ESAT-6 sequences into a DNA vaccine vector and were used to immunize mice. Splenic lymphocytes from vaccinated mice were tested for gamma interferon (IFN-γ) and interleukin-10 (IL-10) secretion. Serum antibodies were examined for P71 antigen-specific isotype responses. When stimulated in vitro with purified P71 antigen, splenocytes from the ESAT-6:P71 vaccinates secreted higher levels of IFN-γ and lower levels of IL-10 compared to those of vaccinates receiving the P71 construct alone. Furthermore, the immunoglobulin G2a serum antibody levels were significantly higher in the ESAT-6:P71 vaccinates compared to those of the vaccinates receiving P71 alone. In conclusion, ESAT-6 was shown to enhance antigen-specific type 1 immune responses in BALB/c mice when used in DNA vaccines.

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Faculty Opinions recommendation of Eukaryotic expression plasmid transfer from the intracellular bacterium Listeria monocytogenes to host cells.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1000439.8005 ◽

2001 ◽

Author(s):

C Graham Clark

Keyword(s):

Listeria Monocytogenes ◽

Plasmid Transfer ◽

Eukaryotic Expression ◽

Intracellular Bacterium ◽

Host Cells ◽

Eukaryotic Expression Plasmid ◽

Expression Plasmid

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Antibody response of carp, Cyprinus carpio to the cestode, Bothriocephalus acheilognathi

Parasitology ◽

10.1017/s0031182099004242 ◽

1999 ◽

Vol 118 (6) ◽

pp. 635-639 ◽

Cited By ~ 11

Author(s):

P. NIE ◽

D. HOOLE

Keyword(s):

Antibody Response ◽

Cyprinus Carpio ◽

Serum Antibody ◽

Infected Fish ◽

Immune Protection ◽

Carp Cyprinus Carpio ◽

Bothriocephalus Acheilognathi ◽

Protection Against Infection ◽

Antibody Levels ◽

Antibody Secreting Cells

The humoral antibody response and the number of pronephric antibody-secreting cells were examined in naturally Bothriocephalus acheilognathi-infected carp. Cyprinus carpio, and in those injected intraperitoneally with an extract of the cestode. In the extract-injected fish, specific antibody was detected 3 weeks after a second injection given 2 weeks after the primary injection, and antibody levels persisted for more than 200 days. A third injection also enhanced the antibody level in the extract-injected carp. The numbers of antibody-secreting cells were significantly higher in carp injected 3 times with the extract than in the control. In naturally-infected fish, the serum antibody levels and the number of pronephric antibody-secreting cells were higher in infected fish than in uninfected individuals although this difference was not statistically significant. The relevance of these results to immune protection against infection is discussed.

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The Caspase-1/IL-18 Axis of the Inflammasome in Tumor Cells: A Modulator of the Th1/Tc1 Response of Tumor-Infiltrating T Lymphocytes in Colorectal Cancer

Cancers ◽

10.3390/cancers13020189 ◽

2021 ◽

Vol 13 (2) ◽

pp. 189

Author(s):

Linda Bilonda Mutala ◽

Cécile Deleine ◽

Matilde Karakachoff ◽

Delphine Dansette ◽

Kathleen Ducoin ◽

...

Keyword(s):

Colorectal Cancer ◽

T Lymphocytes ◽

Tumor Cells ◽

Ex Vivo ◽

Explant Culture ◽

T Cell Response ◽

Mrna Levels ◽

Innate And Adaptive Immunity ◽

Caspase 1 ◽

Culture Model

In colorectal cancer (CRC), a high density of T lymphocytes represents a strong prognostic marker in subtypes of CRC. Optimized immunotherapy strategies to boost this T-cell response are still needed. A good candidate is the inflammasome pathway, an emerging player in cancer immunology that bridges innate and adaptive immunity. Its effector protein caspase-1 matures IL-18 that can promote a T-helper/cytotoxic (Th1/Tc1) response. It is still unknown whether tumor cells from CRC possess a functional caspase-1/IL-18 axis that could modulate the Th1/Tc1 response. We used two independent cohorts of CRC patients to assess IL-18 and caspase-1 expression by tumor cells in relation to the density of TILs and the microsatellite status of CRC. Functional and multiparametric approaches at the protein and mRNA levels were performed on an ex vivo CRC explant culture model. We show that, in the majority of CRCs, tumor cells display an activated and functional caspase-1/IL-18 axis that contributes to drive a Th1/Tc1 response elicited by TILs expressing IL-18Rα. Furthermore, unsupervised clustering identified three clusters of CRCs according to the caspase-1/IL-18/TIL density/interferon gamma (IFNγ) axis and microsatellite status. Together, our results strongly suggest that targeting the caspase-1/IL-18 axis can improve the anti-tumor immune response in subgroups of CRC.

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Non-Lethal Sequential Individual Monitoring of Viremia in Relation to DNA Vaccination in Fish–Example Using a Salmon Alphavirus DNA Vaccine in Atlantic Salmon Salmo salar

Vaccines ◽

10.3390/vaccines9020163 ◽

2021 ◽

Vol 9 (2) ◽

pp. 163

Author(s):

Catherine Collins ◽

Katherine Lester ◽

Jorge Del-Pozo ◽

Bertrand Collet

Keyword(s):

Dna Vaccine ◽

Experimental Infection ◽

Vaccine Efficacy ◽

Dna Vaccines ◽

Experimental Studies ◽

Dna Vaccination ◽

Line System ◽

Individual Fish ◽

Antibody Levels ◽

Subtype 3

Traditionally, commercial testing for vaccine efficacy has relied on the mass infection of vaccinated and unvaccinated animals and the comparison of mortality prevalence and incidence. For some infection models where disease does not cause mortality this approach to testing vaccine efficacy is not useful. Additionally, in fish experimental studies on vaccine efficacy and immune response the norm is that several individuals are lethally sampled at sequential timepoints, and results are extrapolated to represent the kinetics of immune and disease parameters of an individual fish over the entire experimental infection period. In the present study we developed a new approach to vaccine testing for viremic viruses in fish by following the same individuals over the course of a DNA vaccination and experimental infection through repeated blood collection and analyses. Injectable DNA vaccines are particularly efficient against viral disease in fish. To date, two DNA vaccines have been authorised for use in fish farming, one in Canada against Infectious Haemorrhagic Necrotic virus and more recently one in Europe against Salmon Pancreatic Disease virus (SPDv) subtype 3. In the current study we engineered and used an experimental DNA vaccine against SPDv subtype 1. We measured viremia using a reporter cell line system and demonstrated that the viremia phase was completely extinguished following DNA vaccination. Differences in viremia infection kinetics between fish in the placebo group could be related to subsequent antibody levels in the individual fish, with higher antibody levels at terminal sampling in fish showing earlier viremia peaks. The results indicate that sequential non-lethal sampling can highlight associations between infection traits and immune responses measured at asynchronous timepoints and, can provide biological explanations for variation in data. Similar to results observed for the SPDv subtype 3 DNA vaccine, the SPDv subtype 1 DNA vaccine also induced an interferon type 1 response after vaccination and provided high protection against SPDv under laboratory conditions when fish were challenged at 7 weeks post-vaccination.

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Low Immunogenicity of Intravesical Phage Therapy for Urogenitary Tract Infections

Antibiotics ◽

10.3390/antibiotics10060627 ◽

2021 ◽

Vol 10 (6) ◽

pp. 627

Author(s):

Sławomir Letkiewicz ◽

Marzanna Łusiak-Szelachowska ◽

Ryszard Międzybrodzki ◽

Maciej Żaczek ◽

Beata Weber-Dąbrowska ◽

...

Keyword(s):

Clinical Trial ◽

Bacterial Infections ◽

Phage Therapy ◽

Serum Antibody ◽

Multidrug Resistant ◽

Antibody Responses ◽

Urogenital Infections ◽

Reliable Model ◽

Tract Infections ◽

Antibody Levels

Patients with chronic urinary and urogenital multidrug resistant bacterial infections received phage therapy (PT) using intravesical or intravesical and intravaginal phage administration. A single course of PT did not induce significant serum antibody responses against administered phage. Whilst the second cycle of PT caused a significant increase in antibody levels, they nevertheless remained quite low. These data combined with good therapy results achieved in some patients suggest that this mode of PT may be an efficient means of therapy for urogenital infections and a reliable model for a clinical trial of PT.

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