The world’s first malaria vaccine: Hope and challenge / 中国血吸虫病防治杂志

The RTS,S/AS01 is a subunit malaria vaccine against the pre-erythrocytic stage of Plasmodium falciparum. After over 30 years of research and development and clinical trials, this vaccine has been recommended by the WHO for use among children living in highly malaria endemic areas. Although the RTS, S/AS01 vaccine suffers from problems of a low protective efficacy (about 30%), need of four doses and short duration of protective immunity, this malaria vaccine is expected to save tens of thousands of children’s lives, and avoid tens of millions of malaria cases annually, because there have been tens of thousands of childhood deaths due to malaria recently. The introduction of the RTS, S/AS01 vaccine is therefore, widely accepted as a milestone in the history of battle against malaria, which brings a hope to contain malaria and even eventually eliminate malaria. Although there are still multiple challenges in the development of a satisfactory malaria vaccine, the success of the RTS, S/AS01 malaria greatly facilitates the progress towards the development of parasitic disease vaccines, and a more perfect malaria vaccine deserves expectations.

Research progress in Neisseria gonorrhoeae lipooligosaccharides vaccine / 中华微生物学和免疫学杂志

Gonorrhea, caused by Neisseria gonorrhoeae, is one of the most frequently reported infectious diseases. With the increasing antibiotic resistance in Neisseria gonorrhoeae, gonorrhea has become a major public health problem worldwide, making it imperative to develop a safe and effective vaccine. Lipooligosaccharides (LOS), which exist on the outer surface of gram-negative bacteria, contain many important antigenic determinants. In recent years, a large number of studies have shown that LOS may become the most potential target of Neisseria gonorrhoeae vaccine and immunotherapy. This article reviewed the structure of LOS, its role in Neisseria gonorrhoeae infection, research progress in LOS vaccine and the challenges faced in vaccine development, aiming to provide reference for further study.

Protective effects of bicistronic DNA vaccines of herpes simplex virus type 2 glycoprotein D and adjuvant CCL28 on mice / 中华微生物学和免疫学杂志

Objective:To study the protective effects of bicistronic DNA vaccines carrying herpes simplex virus type 2 glycoprotein D (HSV-2 gD) and adjuvant CCL28 sequences that were connected by internal ribosome entry site (IRES) sequence in mouse model.Methods:The recombinant DNA vaccines, pgD-IRES-CCL28 and pCCL28-IRES-gD, encoding HSV-2 gD and adjuvant CCL28 were constructed with IRES sequence. After verified by sequencing, they were intramuscularly injected twice into BALB/c mice. Serum samples and vaginal lavage fluids were collected regularly. Splenocytes, mesenteric lymph node cells and rectal mucosa tissues were separated and collected. The titers of antigen-specific antibodies in immunized mice were analyzed with end-point ELISA. In vitro neutralization assay was used to measure neutralizing antibody titers in serum and vaginal lavage fluid after vaccination and virus challenge. CCL28-responsive immune cells in splenocytes, mesenteric lymph node cells and rectal tissues were detected by chemotaxis experiment and immunohistochemical staining. The protective effects of the bicistronic DNA vaccines were evaluated by fluorescent quantitative PCR, weighing and disease severity assessment. Humoral and cellular immune responses induced by the bicistronic DNA vaccines and their efficacy in immunoprotection were analyzed by comparing with pgD+ pCCL28 group. Results:IgG titers in serum samples and IgA antibody titers in vaginal lavage fluids of mice immunized with pCCL28-IRES-gD were similar to those in pgD+ pCCL28 group. The neutralizing ability of antibodies, the number of rectal mucosal IgA+ plasma cells and CCL28-responsive immune cells in mucosal tissues were increased in pCCL28-IRES-gD group. Serum neutralizing antibodies were not produced immediately in the mice challenged with HSV-2, but no weight loss, disease symptoms or death was observed. However, pgD+ pcDNA3.1 and pgD-IRES-CCL28 were ineffective against HSV-2 infection in mice.Conclusions:The recombinant bicistronic DNA vaccine of pCCL28-IRES-gD could induce stronger mucosal immune response in mice and provide better protective effects.

Protective effects of fusion recombinant DNA vaccine of herpes simplex virus type 2 glycoprotein D and adjuvant CCL19 on mice / 中华微生物学和免疫学杂志

Objective:To compare the immune responses to simply mixed and fused recombinant DNA vaccines of herpes simplex virus type 2 glycoprotein D (HSV-2 gD) and molecular adjuvant CCL19 in mice and to evaluate the protective effects.Methods:Gene recombination technology was used to construct recombinant DNA vaccines expressing HSV-2 gD and CCL19 alone or fused together. After verification by sequencing, Western blot and ELISA, BALB/c mice were immunized twice by intramuscular injection. Serum samples and vaginal lavage fluids were collected regularly after immunization. Splenocytes, mesenteric lymph node cells and rectal tissues were collected after immunization. Differences in humoral and cellular immune responses to the two forms of vaccines and their protective effects in mice were analyzed using end-point ELISA, in vitro neutralization assay, immunohistochemical staining, chemotaxis assay, vaginal virus challenge, fluorescence quantitative PCR, weighing and disease severity assessment. Results:The fused recombinant pgD-IZ-CCL19 plasmid could express gD protein and CCL19 protein in vitro, but the level of expressed CCL19 protein by pCCL19-IZ-gD plasmid was less than that by pgD-IZ-CCL19. The mice immunized with pgD-IZ-CCL19 showed higher levels of IgG in sera and IgA in vaginal lavage fluids ( P<0.01) and stronger neutralization ability than the mice vaccinated with pgD+ pCCL19. Compared with other groups, more lymphocytes were recruited in the rectal mucosa, the spleen and mesenteric lymph nodes of mice immunized with pgD-IZ-CCL19. Weight loss or disease symptoms were not observed in the pgD-IZ-CCL19 group after virus challenge. In addition, the positive rate of HSV-2 in vaginal mucosa and the mortality rate in the pgD-IZ-CCL19 group were the lowest. However, pCCL19-IZ-gD turned out ineffective in preventing HSV-2 infection. Conclusions:The fused recombinant DNA vaccine pgD-IZ-CCL19 could induce stronger immune responses in mice and provide better protective effects, which was superior to the simply mixed DNA vaccine.

Protection against human metapneumovirus (hMPV) conveyed by influenza virus vectors carrying multiple epitope antigens of hMPV / 中华微生物学和免疫学杂志

Objective@#To evaluate the immune responses and protection against human metapneumovirus (hMPV) conveyed by influenza virus vectors carrying multiple epitope antigens of hMPV.@*Methods@#Two recombinant influenza viruses (rFLU/hMPV/B and rFLU/hMPV/CTL+ Th) carrying hMPV multi-epitope gene segments in NS gene were generated by reverse genetic techniques of eight-plasmid system. BALB/c mice were immunized intranasally with rFLU/hMPV/B and rFLU/hMPV/CTL+ Th twice at a two-week interval. Virus-specific antibody titers and splenocyte cytokines were detected two weeks after the boost immunization. Viral loads in lung tissues and turbinates were detected with digital PCR after the immunized mice were challenged with hMPV and influenza virus. Moreover, HE staining was used to observe lung injuries.@*Results@#Specific antibodies against both the influenza virus and hMPV were induced in mice immunized intranasally with rFLU/hMPV/B, while the influenza virus-specific antibody response and hMPV-specific cytotoxic lymphocyte response (significant IFN-γ secretion) were detected in mice immunized with rFLU/hMPV/CTL+ Th. Additionally, balanced Th1/Th2 responses were elicited by rFLU/hMPV/B and rFLU/hMPV/CTL+ Th. Both rFLU/hMPV/B and rFLU/hMPV/CTL+ Th conveyed effective protection against subsequent influenza virus and hMPV challenges with significantly alleviated histopathological damages and reduced viral loads.@*Conclusions@#Both rFLU/hMPV/B and rFLU/hMPV/CTL+ Th can induce specific humoral immune response against hMPV and/or the influenza virus. Moreover, rFLU/hMPV/CTL+ Th can also elicit hMPV-specific CTL immune response. These two recombinant strains can also protect BALB/c mice from the challenges with hMPV and influenza virus, suggesting that they are promising vaccine candidates.

Protection against human metapneumovirus ( hMPV) conveyed by influenza virus vectors carrying multiple epitope antigens of hMPV / 中华微生物学和免疫学杂志

Objective To evaluate the immune responses and protection against human metapneu-movirus ( hMPV) conveyed by influenza virus vectors carrying multiple epitope antigens of hMPV. Methods Two recombinant influenza viruses ( rFLU/hMPV/B and rFLU/hMPV/CTL+Th ) carrying hMPV multi-epitope gene segments in NS gene were generated by reverse genetic techniques of eight-plasmid system. BALB/c mice were immunized intranasally with rFLU/hMPV/B and rFLU/hMPV/CTL+Th twice at a two-week interval. Virus-specific antibody titers and splenocyte cytokines were detected two weeks after the boost immunization. Viral loads in lung tissues and turbinates were detected with digital PCR after the immunized mice were challenged with hMPV and influenza virus. Moreover, HE staining was used to observe lung inju-ries. Results Specific antibodies against both the influenza virus and hMPV were induced in mice immu-nized intranasally with rFLU/hMPV/B, while the influenza virus-specific antibody response and hMPV-spe-cific cytotoxic lymphocyte response ( significant IFN-γ secretion ) were detected in mice immunized with rFLU/hMPV/CTL+Th. Additionally, balanced Th1/Th2 responses were elicited by rFLU/hMPV/B and rFLU/hMPV/CTL+Th. Both rFLU/hMPV/B and rFLU/hMPV/CTL+Th conveyed effective protection against subsequent influenza virus and hMPV challenges with significantly alleviated histopathological dama-ges and reduced viral loads. Conclusions Both rFLU/hMPV/B and rFLU/hMPV/CTL+Th can induce spe-cific humoral immune response against hMPV and/or the influenza virus. Moreover, rFLU/hMPV/CTL+Th can also elicit hMPV-specific CTL immune response. These two recombinant strains can also protect BALB/c mice from the challenges with hMPV and influenza virus, suggesting that they are promising vaccine candi-dates.

Comparison of immunoadjuvant activities of four bursal peptides combined with H9N2 avian influenza virus vaccine

The bursa of Fabricius (BF) is a central humoral immune organ unique to birds. Four bursal peptides (BP-I, BP-II, BP-III, and BP-IV) have been isolated and identified from the BF. In this study, the immunoadjuvant activities of BPs I to IV were examined in mice immunized with H9N2 avian influenza virus (AIV) vaccine. The results suggested that BP-I effectively enhanced cell-mediated immune responses, increased the secretion of Th1 (interferon gamma)- and Th2 (interleukin-4)-type cytokines, and induced an improved cytotoxic T-lymphocyte (CTL) response to the H9N2 virus. BP-II mainly elevated specific antibody production, especially neutralizing antibodies, and increased Th1- and Th2-type cytokine secretion. BP-III had no significant effect on antibody production or cell-mediated immune responses compared to those in the control group. A strong immune response at both the humoral and cellular levels was induced by BP-IV. Furthermore, a virus challenge experiment followed by H&E staining revealed that BP-I and BP-II promoted removal of the virus and conferred protection in mouse lungs. BP-IV significantly reduced viral titers and histopathological changes and contributed to protection against H9N2 AIV challenge in mouse lungs. This study further elucidated the immunoadjuvant activities of BPs I to IV, providing a novel insight into immunoadjuvants for use in vaccine design.

Advances in prevention and control of yellow fever / 中国人兽共患病学报

Yellow fever(YF) is a natural focal disease transmitted by infected mosquitoes,which had caused epidemic in some areas of Europe and America port city in the history,bringing great disaster to human society.Since 40s of last century,a series of derivative strain of 17D vaccines,providing lifelong protection,had been vaccinated a large population who are trave ling to or living in areas at risk for YF,which avoid the outbreak of the disease,appearing quiet state.Since the beginning of 1990s,the disease had outbreak successively in Africa,tropical American areas and its nations.The mortality was even more higher than Ebola happened in 3 countries in West Africa between 2014 and 2015.In 2016-2017the large outbreak in Angola,Congo and Brazil,has claimed thousands of lives.Yellow fever has never occurred in Asia until the introduction of 11 cases by jet travel from Angola to China,which attracted the attention of Chinese government.Yellow fever elimination initiative strategic plan for 2017 2026 was launched by WHO,mainly based on that 17D vaccine which is safe and effective to control the spread of this disease.The measures of controlling mosquitoes and vaccination programs,in particular putting great expectations for the latter,are the main measures for prevention the disease and achievement the goal of WHO elimination strategic plan.But strong guarantees of related countries and abiding by the provisions of the global quarantine regulations are also important.Therefore,this article reviews the historical and the recent epidemic outbreaks of yellow fever,vaccination and other prevention measures,hoping that the WHO elimination of the yellow fever epidemic strategy will achieve successfully in 2026.

Clinical Observation of Bevacizumab Combined with FOLFIRI Plan in the Treatment of Advanced Colon Cancer / 中国药房

OBJECTIVE:To observe therapentic efficacy of bevacizumab combined with irinotecan+leucovorin+fluorouracil (FOLFIRI)plan in the treatment of advanced colon cancer,toxic reaction and patients'survival rate. METHODS:A total of 113 patients with advanced colon cancer admitted to the oncology department in our hospital from Jan. 2010 to Aug. 2014 were random-ized into observation group 1(40 cases),observation group 2(39 cases)and control group(34 cases). Three groups received FOLFIRI;observation group 1 and 2 were additionally given Bevacizumab injection 5 and 7.5 mg/kg 14 d as a treatment course, for 8 cycles. Clinical efficaices as well as the positive rate of VEGF-A,immune indexes(the proportion of CD3+,CD3+CD4+,CD3+CD8+ in T cell subset)before and after treatment,the incidence of toxic reaction,1-year and 2-year survival rates were compared among 3 groups. RESULTS:The total response rate of observation group 1 and 2 were significantly higher than control group,and the positive rate of VEGF-A in observation group 1 and 2 were significantly lower than control group,with statistical significance (P0.05). 2-year survival rate of observation group 1 and 2 were significantly higher than control group,with statistical significance(P<0.05). CONCLUSIONS:For advanced colon cancer,different doses of bevacizumab combined with FOLFIRI have significant synergistic effect,can effectively inhibit VE-FG-A,play a role of immune protection and anti-toxic side effects,and prolong the survival time. The incidence of hypertension in patients treated with low-dose bevacizumab is relatively lower and the safety is better.

Protective effect of interferon-γ on Chlamydia psittaci acute infection / 中国人兽共患病学报

We investigated the effects of IFN-γ on Chlamydia psittaci (Cps) infection.HeLa cells were treated with different concentrations of recombinant human IFN-γ (5 ng/mL,25 ng/mL,50 ng/mL) after infecting with C.psittaci 6BC,then the number and morphology of C.psittaci inclusion bodies were examined after 48 hours.C57BL/6J mice were intranasally infected with 2 × 106 IFUs C.psittaci 6BC,and intraperitoneally administrated with 10 μg recombinant murine interferon-γ 24 hours prior or post infection,then body weight,activity and survival rate were recorded.The histopathology of mice livers and lungs was analyzed by HE staining on day 5 or day10 post infection.And the chlamydial inclusion bodies were titrated in the lung homogenates of mice sacrificed on day 5 after infection.The inclusion body numbers of recombinant human IFN-γ treated groups (by 5ng/mL,25ng/mL,50ng/mL) were significantly less than that in the control group (23.8±5.1)× 106,(10± 3.58) × 106,(8.0±2.22) × 106,(43.3±11.05)× 106,respectively).And the morphology of inclusion bodies in IFN-γ treated HeLa cells was irregular and much smaller.We also found that IFN-γ could significantly improve the survival rate,reduce acute clinical manifestations and pathological injurery of lung and liver in C.psittaci respiratory tract infected mice model.So we summarized that IFN-γ can mediate strong immunological protection during acute C.psittaci early infection.

Preliminary research on prokaryotic expression and immune protection of triosephosphate isomerase of Toxoplasma gondii / 中国血吸虫病防治杂志

Objective To study the prokaryotic expression and immune protection of triosephosphate isomerase(TPI)of Toxoplasma gondii in mice. Methods Total RNA was extracted from toxoplasma tachyzoites,and TPI fragment was amplified by PCR and cloned into the prokaryotic expression vector pET-28a(+). The target protein was induced with IPTG and purified by Ni-NTA affinity chromatography. The mice were immunized 4 times by emulsified TPI with adjuvant,and the last time was the strengthen immunization. At the same time,an adjuvant group and a normal group were set as controls. The blood samples were got from the tail vein of the mice,and the serum antibody titres were detected. All the mice were challenged with 400 toxo-plasma tachyzoites to observe the survival time. Results The TPI gene was amplified from T. gondii cDNA by PCR. The recom-binant vector TPI/pET-28a(+)was usefully constructed,and the TPI protein was expressed and purified. The serum antibody ti-tre could be more than 100 thousand. After infected with toxoplasma tachyzoites,the survival time of the mice in the experimen-tal group was longer than that of the mice in the control groups. Conclusion The TPI protein of T. gondii could trigger the im-munoprotection against T. gondii challenge in the mice.

Immune protective effect of recombinant growth hormone in chronic obstructive pulmonary disease complicated with respiratory failure treated by ventilator therapy / 中国生化药物杂志

Objective To investigate the immune protective effect of recombinant growth hormone on chronic obstructive pulmonary disease ( COPD) patients with typeⅡin respiratory failure during the application of ventilator.Methods 52 patients with COPD with respiratory failure in Taizhou City Hospital from July 2014 to November 2015 in this study who were treated with ventilator therapy were divided into two groups.The control group received routine treatment, and the treatment group received more with recombinant growth hormone 8 U/d subcutaneous injected.Before and after treatment 14 days,determination of nutritional indicators,immune function,inflammatory markers and indicators of oxidative stress.Results Compared with before treatment,levels of total protein,albumin,transferrin and fibronectin in 2 groups after treatment increased,levels of IgM,IgG,CD4 +,CD8 +and CD4 +/CD8 +increased,levels of TNF-α,IL-8 and CRP decreased,levels of MDA,GSH and TAO decreased,levels of SOD increased(P<0.05), compared with the control group,levels of total protein,albumin,transferrin and fibronectin in the treatment group were higher,levels of IgM,IgG,CD4 +, CD8 +and CD4 +/CD8 +were higher,levels of TNF-α,IL-8 and CRP were lower,levels of MDA,GSH and TAO were lower,levels of SOD were higher (P<0.05).Conclusion Recombinant growth hormone has immune function protective effect on the COPD patients with type II respiratory failure in the process of breathing machine, which can improve the nutritional index and reduce the inflammatory reaction.

Evaluation of immune protection by inactivated whole bacteria vaccine of Stenotro-phomonas maltophilia / 军事医学

Objective To evaluate the immunoprotection by the inactivated whole bacteria(IWB) of Stenotrophomonas maltophilia K279a in mice.Methods Mice were immunized by inactivated whole bacteria of S.maltophilia K279a made from formaldehyde.When the indicated the antibody titer of the mice reached the require level, the protective effect of the IWB was evaluated by performing the opsonophagocytic killing test in vitro and the poison attack experiments in vivo. Results It was found that IgG in serum of the immunized mice measured by ELISA was significantly increased after the second immune enhancement, and antiserum in vitro had strong phagocytic effect.Meanwhile, immunoprotection of the immunized groups was also significantly increased when challenged by S.maltophilia K279a.Conclusion Effective humoral immune response can be predominantly induced by the inactivated whole bacteria of S.maltophilia K279a, providing protection against challenge by S.maltophilia K279a in BALB/c mice.

Inmunoprotección activa contra cocaína / Active immunoprotection to cocaine

INTRODUCCIÓN: La farmacopea clásica, empleada para atenuar la dependencia a ciertas drogas de abuso ilegal, como la cocaína, ha demostrado una pobre eficacia terapéutica. Basado en este desalentador panorama clínico-terapéutico, desde hace más de una década diversos investigadores han desarrollado nuevas estrategias terapéuticas contra la adicción a la cocaína. Estas nuevas estrategias experimentales están basadas en el diseño y la síntesis de formulaciones estructurales de vacunas terapéuticas contra la adicción a la cocaína. OBJETIVO: Realizar una descripción del desarrollo y la validación terapéutica de la inmunización activa contra la cocaína. MÉTODO: Se realizó una búsqueda bibliográfica con el uso del PubMed, usando como descriptores las palabras "Cocaine" y "Vaccine". Se obtuvieron 155 artículos, de los cuales se usaron 46 para esta revisión. RESULTADOS: A nivel preclínico, la vacunación activa genera altos niveles de anticuerpos capaces de reconocer con alta especificidad a la cocaína dentro del torrente sanguíneo, atenuando las alteraciones conductuales inducidas por diversas dosis de cocaína. DISCUSIÓN Y CONCLUSIÓN: Los resultados preclínicos y clínicos han reforzado "la prueba de concepto" terapéutica de la vacunación activa para el control farmacológico de la recaída al consumo adictivo de la cocaína en el humano, sin embargo, dieron pauta a la postulación y a la justificación de sintetizar nuevos modelos de uso humano de vacunas anticocaína. Esta estrategia farmacológica experimental, de naturaleza "inmunoprotectora", ha demostrado ser un tratamiento eficaz al atenuar significativamente las conductas de búsqueda y consumo adictivo a la cocaína, tanto a nivel pre-clínico, en el modelo del roedor, como en el humano.

INTRODUCTION: The classic pharmacopoeia used to attenuate cocaine dependence has proved a poor therapeutic efficacy. Based on this discouraging clinical and therapeutic panorama, since more than a decade, various researchers have developed new therapeutic strategies against cocaine addiction. These new experimental strategies are based on the structural design and synthesis of therapeutic vaccine formulations against cocaine addiction. OBJECTIVE: To describe the development and therapeutic evaluation of active immunization against cocaine. METHOD: A bibliographical search was made using PubMed, using as descriptors the words "Cocaine" and "Vaccine." 155 articles were obtained which were used for these review 46 items. RESULTS: At preclinical level, active vaccination generates high levels of antibodies capable of recognizing with high specificity the cocaine present in the bloodstream, which attenuates the behavioral changes induced by different doses of cocaine. DISCUSSION AND CONCLUSION: Preclinical and clinical results have reinforced "proof of concept" active therapeutic vaccination to pharmacological control to cocaine use relapse in humans, but gave guidelines to the postulation and justification of synthesizing new models of anti-cocaine vaccines for human use. This experimental pharmacological strategy of "immunoprotective" nature has proven an effective treatment that significantly reduces drug-seeking behaviors, both at pre-clinical levels in the rodent model as well as in humans.

Cloning and function analysis of high mobility group box 1(HMGB1)protein of Schistosoma japonicum(Mainland strain) / 中国血吸虫病防治杂志

Objective To clone and express a high mobility group box 1(HMGB1)protein of Schistosoma japonicum(Main-land strain)and analyze its function. Methods The DNA fragment of open reading frame encoding Sj HMGB1 protein was ampli-fied by RT-PCR from the mRNA of S. japonicum worms,then it was subcloned into the expression vector pET28a(+)to form the recombinant expression plasmid SjHMGB1-pET28a. The recombinant expression plasmid was transformed into the component E. coli BL21(DE3),and the tranformant containing recombinant expression plasmid was induced with IPTG to express the recombi-nant protein SjHMGB1. The recombinant SjHMGB1 protein was purified by affinity chromatography with nickel chelating affinity chromatography agarose gel. The Gel retard experiment and animal immunization were performed to analyze the DNA binding ca- pacity and the immunologic property of recombinant SjHMGB1. The expression levels of HMGB1 in different life cycle stages of S. japonicum were analyzed by Western bloting and RT-PCR. Female ICR mice were immunized with the recombinant SjHMGB1 pro-tein and infected with 45±2 cercariae of S. japonicum after three immunizations. Forty-two days post-infection,the worms and eggs of S. japonicum were recovered from the portal vein and liver tissue,respectively. The worm and egg reduction rates were calculat-ed respectively. Results A 530 bp of specific DNA fragment was amplified from mRNA of S. japonicum by RT-PCR,which was the open reading frame(ORF)encoding SjHMGB1protein confirmed by DNA sequencing analysis. The recombinant expression plasmid SjHMGB1-pET28a was constructed by cloning the ORF of SjHMGB1 into a expression vector pET28a(+). The bacterium transformants containing the recombinant plasmid expressed a soluble recombinant protein about 28 kDa after induced by IPTG, and the recombinant SjHMGB1 protein was purified by nickel chelating affinity chromatography. The gel retard experiment showed that the recombinant SjHMGB1 protein could bind to both supercoiled DNA and linear DNA,and the recombinant protein immu-nized mice produced high titers of antiserum IgG. Western bloting indicated that the recombinant SjHMGB1 protein was recognized specifically by the S. japonicum-infected mice serum. Above results showed that the recombinant SjHMGB1 protein possessed both functional activity and immunogenicity as the natural protein. RT-PCR and Western blot results showed that SjHMGB1 was abun-dantly expressed in the adult and egg stages whereas barely detectable in the cercaria stage. The immune protection experiment showed that the recombinant SjHMGB1 induced mice to produce high titers of specific antibody IgG but failed to conduct an effec-tive immune protection against S. japonicum. Conclusion The gene encoding HMGB1 from S. japonicum and the soluble recombi-nant SjHMGB1 protein with natural functional activity are obtained,and the recombinant SjHMGB1 has a high immunogenicity but is not able to induce an effective immune protection against S. japonicum.

The Immune Protective Effect of Astragalus Polysaccharide on Mice Genital Tract Infected with Chlamydia Trachomatis / 浙江中医药大学学报

[Objective]To probe into the immune protective effect of Astragalus polysaccharide on mice ,whose genital tract was infected with Chlamydia trachomatis. [Methods]After the murine model of genital tract infected with chlamydia trachomatis was established, we could obtain the minimum effective infective dose. Mice were immuned by different doses(10 mg·kg-1, 20 mg·kg-1, 40 mg·kg-1) of Astragalus polysaccharide(APS) and 0.9%NS , meanwhile the blank group was regarded as the control group. After mice were immuned, we detected serum IgG antibody levels by ELISA method,and took the twice effective dose of chlamydia trachomatis to attack the genital tract after a week. Then the mice genital shedding of epithelial cells were cultured at each interval 2d. After 10 days, we kil ed the mice to calculate spleen(thymus) index, and observed the pathological changes in the reproductive tract of the mice. [Results]Compared with the control group, spleen(thymus) index and serum IgG antibody levels of Astragalus polysaccharide group are significantly higher. The results of vaginal exfoliated cells from Ct training show that, Ct positive rate of Astragalus polysaccharide group is significantly reduced, the number of inclusion bodies is also significantly decreased. And with the increasing concentration of Astragalus polysaccharide, the protective effect is more obvious. The NS group has no significant difference from the control group in al indicators. [Conclusions]Astragalus polysaccharide can induce immune protection on Ct genital tract attacked mice, and in the concentration range, with the increase of the concentration, the protection effect is better.

Ag85A DNA vaccination boosting enhances BCG primed-mice anti-tuberculosis T cell responses / 中华微生物学和免疫学杂志

Objective To construct DNA vaccine expressing Mycobacterium tuberculosis(Mtb) immunodominant antigen Ag85A and analyze its anti-tuberculosis T cell responses in BCG primed-mice after DNA vaccination boosting.Methods The coding gene of Ag85A mature fragment was amplified by PCR with H37Rv genomic DNA as template,and then cloned into the eukaryotic expression vector pVAX1 to construct Ag85A DNA vaccine.After purification,Ag85A DNA vaccine was injected intramuscularly twice in BCG primed-mice with BCG vaccination and DNA vaccination alone as control.Eight weeks post-vaccination,spleen lymphocytes were separated and were then used to analyze Mtb antigen specific effector T cell response and polyfuntional IFN-γ/TNF-α/IL-2 secreting CD4+ T cell frequencies and intensities,and CD8+T cell responses by IFN-γ ELISPOT assay and intracellular staining,respectively.Results Compared to BCG vaccinated-and DNA vaccinated-mice,Ag85A DNA boosting not only enhanced significantly BCG primed-mice IFN-γ+TNF-α+IL-2+,IFN-γ+ IL-2+,TNF-α+IL-2+ and IL-2+ CD4+ T cell frequencies and IL-2 secretion,but also improved significantly IFN-γ-secreting and IL-2-secreting CD8+ T cell frequencies.Condusion Ag85A DNA vaccine was constructed successfully and was demonstrated to enhance significantly BCG primed-mice Mtb antigen specific CD4+ and CD8+ T cell responses when boosting,which is beneficial to improve BCG immunogenicity and its waning immune protection against Mtb.

Monoclonal auto-antibodies and sera of autoimmune patients react with Plasmodium falciparum and inhibit its in vitro growth

The relationship between autoimmunity and malaria is not well understood. To determine whether autoimmune responses have a protective role during malaria, we studied the pattern of reactivity to plasmodial antigens of sera from 93 patients with 14 different autoimmune diseases (AID) who were not previously exposed to malaria. Sera from patients with 13 different AID reacted against Plasmodium falciparum by indirect fluorescent antibody test with frequencies varying from 33-100 percent. In addition, sera from 37 AID patients were tested for reactivity against Plasmodium yoelii 17XNL and the asexual blood stage forms of three different P. falciparum strains. In general, the frequency of reactive sera was higher against young trophozoites than schizonts (p < 0.05 for 2 strains), indicating that the antigenic determinants targeted by the tested AID sera might be more highly expressed by the former stage. The ability of monoclonal auto-antibodies (auto-Ab) to inhibit P. falciparum growth in vitro was also tested. Thirteen of the 18 monoclonal auto-Ab tested (72 percent), but none of the control monoclonal antibodies, inhibited parasite growth, in some cases by greater than 40 percent. We conclude that autoimmune responses mediated by auto-Ab may present anti-plasmodial activity.

Construction, expression and immunological protection of DNA vaccine of pVAX1-oipA of Helicobacter pylori / 中国免疫学杂志

Objective:To investigate the effect of DNA vaccine expressed Helicobacter pylori(Hp) oipA on protecting against Hp infection.Methods:The ORFs of Hp oipA had been inserted into the eukaryotic expressing vector pVAX1 and SGC-7901 cells had been transfected with recombinant plasmid pVAX1-oipA. The expression of oipA had been detected by RT-PCR and ELISA. After extracted and purified, pVAX1-oipA had been injected into BALB/c mice through muscles of right leg one time each week for three weeks(100 ?g each mouse). pVAX1 blank and normal saline had been used as the control groups. Titer of antibodies had been detected by ELISA two months after the last immunization. Based on the confirmation of immunological response in the pVAX1 groups, mice had been given orogastric challenged with live Hp Sydney strain three times(0.5?108/0.5 ml each mouse). Four weeks after challenge, mice had been sacrificed. Histological change and the colonization of Hp in the gastric mucosa had been detected by urease test, the culture of Hp, and electronic microscopy.Results:SGC-7901 cells transfected with pVAX1-oipA had expressed corresponding production at the level of transcription and translation. Immunized mice had been induced anti-oipA antibodies. After challenged with bacterium, as contrast to immunized mice groups injected with pVAX1-oipA, pVAX1 blank, normal saline, the positive rate of urease test of gastric mucosa was 0(0/10), 90%(9/10), 100%(5/5)respectively,the positive rate of cultures of Hp was 20%(2/10), 90%(9/10), 100%(5/5)respectively. Histological findings: the different degree of erosion had been observed in control group, but 80%(8/10)of gastric mucosa were normal in immunized mice.Conclusion:oipA DNA could induce effective immune response in protection against Hp infection.

Anti-Tumor Efficacy Induced by Antibodized PDTRP Gene Immunization / 中国肿瘤生物治疗杂志

Objective: To study the anti-tumor efficacy induced by antibodized tumor epitope PDTRP gene immunization. Methods: Three copies of tumor associate gene PDTRP from MUCI tandem repeats were designed and mimicked the conformation of MUCI by Insight Ⅱ . The ?lneo-PDTRP plasmid was further constructed, in which the PDTRP target gene was inserted into CDR3 of the ?1 -neo vector. The specific humoral and cellular immune responses towards to PDTRP were detected after intraspleen immunized Balb/c mice with "ylneo-PDTRP. And the immune protection assay was also done to observe whether the mice immunized with ?lneo-PDTRP could prolong the survival after tumor challenge. Results: The conformation of three copies of PDTRP mimicked the conformation of MUCI tandem repeats. The expression of ?lneo-PDTRP could be detected after in vitro transfect. The specific antibody against PDTRP epitope could be induced and increase to a higher titer after intraspleen injection with a ?lneo-PDTRP plasmid. And the specific proliferation and cytotox-ic function of lymphocyte were also increased. There is a significant survival from mice immunized with ?lneo-PDTRP a-gainst the 4T1-PDTRP tumor challenge. Conclusions: Gene immunization with ?lneo-PDTRP could elicit both humoral and cellular tumor specific immune response and had the protective effect.