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1.
实验中对无细胞百日咳疫苗的脱毒工序优化后,采用双抗夹心ELISA法来检测百日咳有效组分含量,同时采用效价试验方法来验证结果。ELISA法定量测定有效成分的结果和效价试验结果均证明达到《中国药典》三部2005版的要求。由于双抗夹心ELISA法特异性强,灵敏度高,适用于无细胞百日咳疫苗生产各个环节的质量控制。 相似文献
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Semenova IB Remova TN Bazhanova IG Britsina MV Zakharova NS 《Zhurnal mikrobiologii, epidemiologii, i immunobiologii》2004,(1):45-49
The immunomodulating activity of acellular pertussis vaccine (APV) and adsorbed DPT vaccine with acellular pertussis component (DPTA vaccine) was studied. The study revealed that only large doses of APV, 10 immunizing doses (ID), suppressed humoral and cell-mediated response to sheep red blood cells (SRBC). 1 ID produced no influence on the formation of antibody producing cells, but increased the development of delayed hypersensitivity (DH) to SRBC. The modulation of cell-mediated immune response, induced by APV, returned to normal after the injection of purified staphylococcal toxoid, used as immunomodulator, in doses of 0.15 BU per mouse and 1.5 BU per mouse. DPTA vaccine containing 1 ID, as well as 10 ID, produced no immunomodulating effect. This was established by the evaluation of humoral response to SRBC in CBA mice and the study of the formation of DH to SRBC in BALB/c mice. As indicated by the total of the presented data, the inclusion of APV into DPTA vaccine enhanced the immunological safety of its pertussis component. 相似文献
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Zhu YZ Cai CS Zhang W Guo HX Zhang JP Ji YY Ma GY Wu JL Li QT Lu CP Guo XK 《PloS one》2010,5(11):e13915
Background
Pertussis (whooping cough) caused by Bordetella pertussis (B.p), continues to be a serious public health threat. Vaccination is the most economical and effective strategy for preventing and controlling pertussis. However, few systematic investigations of actual human immune responses to pertussis vaccines have been performed. Therefore, we utilized a combination of two-dimensional electrophoresis (2-DE), immunoblotting, and mass spectrometry to reveal the entire antigenic proteome of whole-cell pertussis vaccine (WCV) targeted by the human immune system as a first step toward evaluating the repertoire of human humoral immune responses against WCV.Methodology/Principal Findings
Immunoproteomic profiling of total membrane enriched proteins and extracellular proteins of Chinese WCV strain 58003 identified a total of 30 immunoreactive proteins. Seven are known pertussis antigens including Pertactin, Serum resistance protein, chaperonin GroEL and two OMP porins. Sixteen have been documented to be immunogenic in other pathogens but not in B.p, and the immunogenicity of the last seven proteins was found for the first time. Furthermore, by comparison of the human and murine immunoproteomes of B.p, with the exception of four human immunoreactive proteins that were also reactive with mouse immune sera, a unique group of antigens including more than 20 novel immunoreactive proteins that uniquely reacted with human immune serum was confirmed.Conclusions/Significance
This study is the first time that the repertoire of human serum antibody responses against WCV was comprehensively investigated, and a small number of previously unidentified antigens of WCV were also found by means of the classic immunoproteomic strategy. Further research on these newly identified predominant antigens of B.p exclusively against humans will not only remarkably accelerate the development of diagnostic biomarkers and subunit vaccines but also provide detailed insight into human immunity mechanisms against WCV. In particular, this work highlights the heterogeneity of the B.p immunoreactivity patterns of the mouse model and the human host. 相似文献4.
Zakharova NS Britsina MV Ozeretskovskaia MN Bazhanova IG Mertsalova NU Poddubikov AV Zaĭtsev EM Ermolova EV 《Zhurnal mikrobiologii, epidemiologii, i immunobiologii》2006,(2):46-49
The introduction of the immunomodulator polyoxidonium in an amount of 0.5 Mg/ml into adsorbed D(a)PT vaccine with the acellular pertussis component leads to the preservation of the protective activity of the pertusis component, diphtheria and tetanus toxoids, as well to the 4-time decrease of the content of adsorbent (aluminium hydroxide) from 2 to 0.5 mg/ml. 相似文献
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Gzyl A Augustynowicz E Zawadka M Rabczenko D Slusarczyk J 《Medycyna do?wiadczalna i mikrobiologia》2007,59(2):137-147
Studies concerned evaluation of differences between parameters of cell-mediated immunity in mice, induced with whole-cell and acellular pertussis vaccines with subsequent challenge with B. pertussis strains harbouring different ptxS1/prn allele genes. In the study, concentrations of IFN-gamma/Il-2 and 1l-4/Il-5 in supernatants of cultured mice splenocytes have been determined to evaluate differences in Th1 or Th2 lymphocytes subpopulation response. Simultaneously, studies of intracellular expression of genes encoding of Il-2, Il-12, IFN-gamma and Il-4, Il-5, Il-10, Il-13 in mice splenocytes, and genes encoding factors involved in inflammatory process in the lung tissue (GM-CSF, TNF-alpha, Il-1beta, Il-6 i TGF-beta) have been performed on RNA level. The obtained results, confirmed high polarization of immunological response toward Th1 in mice immunized with DTP vaccine with whole-cell pertussis component, and toward Th2 in mice immunized with acellular pertussis vaccine. Inflammatory process in the lung tissue was more pronounced in animals immunized with whole-cell pertussis vaccine. There were no quantitative differences of analysed factors involved in the immune response among mice challenged B. pertussis strains containing different ptxS1/prn composition. 相似文献
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Anukam Kingsley C. Osazuwa Emmanuel E. Mbata Theodore I. Ahonkhai Ijeoma N. 《World journal of microbiology & biotechnology》2004,20(3):231-234
We investigated 296 adolescents (11–18 years), who had been immunized previously with the three doses of DPT vaccines. 48
were diagnosed positive for HIV-1. Nasopharyngeal swabs were obtained from 296 adolescents who presented with persistent cough
and nasopharyngeal secretions. Nasopharyngeal swabs (calcium alginate) specimens were collected by passing the swabs through
the nares into the posterior nasopharynx and rotating the swabs for a few seconds. The swabs were plated for culture of Bordetella organisms in charcoal cephalexin blood agar (CCBA). The CCBA plates were incubated for 2–6 days at 35 °C in a humid aerobic
atmosphere. The suspected, shiny (mercury-like) colonies were tested by slide agglutination with antisera to B. pertussis and B. parapertussis, and urease, oxidase activities were performed. Results indicate that out of 48 HIV-1-positive adolescents, 18 had positive
cultures for Bordetella organisms (14, Bordetella pertussis, and 4, Bordetella parapertussis). Of 248 HIV-1-negative subjects, 3 had Bordetella organisms (2, Bordetella pertussis, 1, Bordetella bronchiseptica). One of the subjects, a boy, aged 14 years, with Bordetella bronchiseptica had a dog as pet, which was found to be infected. The results indicate that adolescents with HIV-1 infection, despite being
vaccinated against pertussis have a higher rate of infection when exposed to pertussis bacteria than HIV-1-negative adolescents.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
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Zakharova NS Agal'tsova SI Shmeleva EI Britsina MV Ozeretskovskaia MN Remova TN Bazhanova IG Mertsalova NU Ermolova EV 《Zhurnal mikrobiologii, epidemiologii, i immunobiologii》2003,(3):30-34
Toxic properties of acellular pertussis vaccine (APV) and morphological changes in white mice in response to intramuscular injection of APV (without or with immunomodulator glucosaminylmuramyl dipeptide-GMDP) were under study. APV used in these experiments was developed at the Mechnikov Research Institute for Vaccines and Sera (the Russian Acad. Med. Sci.) on the basis of Bordetella pertussis cultures in synthetic fluid culture media. In experiments on acute and chronic toxicity of APV (without GMDP) increased tissue immunity reactions in spleen, thymus, liver, lungs and intestinal wall was detected. There was no difference in immunomorphological reactions in mice receiving APV with different doses of GMDP, but some difference was observed in time dynamics of tissue immunity reactions. A small dose of GMDP should be preferred (0.0001 microgram) which results in gradual growth of tissue immunity reactions less pronounced toxic reactions caused be the APV injection. 相似文献
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The antibody response to filamentous haemagglutinin and pertussis toxin was studied in N:NIH mice vaccinated according to the WHO recommendations for potency test with the International Standard for Pertussis Vaccine (ISPV). Some of the vaccinated animals were challenged intracerebrally on day 14. All animals, whether challenged or not, were bled on days 7, 14, 21, 28 and 35 after immunization. The relationship between anti-PT and anti-FHA antibodies measured by ELISA and protection from intracerebral challenge was examined. All those mice with anti-PT titres on day 14 higher than 43 EU/ml survived challenge. No relationship was found between anti-FHA antibodies and survival. Anti-PT titres on day 14 below 43 EU/ml were related to the days of survival after challenge; a linear regression curve of y = 13 + 2.4x, with a correlation coefficient r = 0.61 was found. Anti-PT antibodies seem to play an important role in protection when animals are challenged intracerebrally, as is the case in the standard potency test for pertussis vaccine. 相似文献
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E J Wiertz H G Loggen H C Walvoort J G Kreeftenberg 《Journal of biological standardization》1989,17(2):181-190
The in vitro response of human B- and T-lymphocytes to the acellular vaccines JNIH-6 (containing pertussis toxoid and filamentous hemagglutinin), and JNIH-7 (containing pertussis toxoid), and to the purified components JNIH-4 (filamentous hemagglutinin) and JNIH-5 (pertussis toxin) was investigated. Pertussis toxoid and filamentous hemagglutinin induced specific Ig synthesis in vitro in lymphocytes obtained from convalescent pertussis patients as target cells. The antigen-dependent Ig production was demonstrated in lymphocyte culture supernatants by ELISA techniques and by a chinese hamster ovary cell toxin neutralization assay. Particularly with JNIH-4, -6 and -7, high antibody titers were obtained. At optimal antigen concentrations a marked lymphocyte blast transformation was found in lymphocyte cultures from whooping cough patients, but not in cultures of lymphocytes obtained from healthy volunteers. At high concentrations native pertussis toxin as well as the B oligomer (S2-5) of the toxin induced a strong proliferation of patient as well as control lymphocytes, indicating non-specific mitogenic activity. At lower concentrations lymphocyte blast transformation was seen in patient cultures only, which indicates an antigen-specific T-cell response. The A protomer (S1), dimer 1 (S2 + 4) and dimer 2 (S3 + 4) induced proliferation of patient lymphocytes, which demonstrates the presence of T-cell epitopes on these peptides. The in vitro B-cell response and the lymphocyte blast transformation assay are both useful tools for estimating the potency of acellular pertussis vaccines in man. Spontaneously acquired and vaccine induced immunity to Bordetella pertussis can be investigated at the level of B- and T-lymphocytes. 相似文献
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M D Priadkina V A Fradkin L S Kuznetsova E K Raetskaia 《Zhurnal mikrobiologii, epidemiologii, i immunobiologii》1975,(3):78-80
The authors present materials on the study of the reaction of blood neutrophils (PPN test) to pertussis allergen in children suffering from pertussis and immunized with ADPT vaccine. Results obtained in examining 111 children showed that the PPN test was specific and could be used for assessment of allergic manifestations in children suffering from pertussis or immunized with ADPT vaccine. Taking into consideration the harmlessness and expressiveness of the PPN test it can be recommended for studying in dynamics in any age groups. 相似文献
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The efficacy of six acellular pertussis vaccines, prepared by various manufacturers in Japan, was investigated in a murine model of respiratory infection (aerosol challenge model) and a murine intracerebral (i.c.) challenge model. There was a good correlation between bacterial clearance from the lungs after aerosol challenge and the potency of vaccines as determined by i.c. challenge. The levels of antibodies against filamentous hemagglutinin were higher after immunizations with all tested vaccines than the levels of antibodies against pertussis toxin and pertactin. Spleen cells from mice immunized with each individual vaccine secreted interferon gamma (IFN-gamma) in response to stimulation by pertussis toxin, filamentous hemagglutinin and fimbriae. The production of interleukin-4 in response to each of the antigens tested was detected but was lower than that of IFN-gamma. However, antibody levels and cell-mediated immune responses were not correlated with the protective effects of the vaccines after aerosol challenge and after i.c. challenge. 相似文献
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伤寒Vi多糖结合疫苗和Vi多糖疫苗分别免疫小鼠,分离血清,采用间接ELISA法测定不同时点血清中特异性IgA、IgM、IgG及其亚类(IgG1、IgG2a、IgG3)的抗体滴度。结果显示,免疫一针后,Vi多糖结合疫苗组的IgG抗体GMT值明显升高,第二针有加强效应(P<0.01);所测3种IgG亚型中IgG2a抗体滴度升高明显;Vi多糖和结合疫苗免疫小鼠后,血清中IgA和IgM抗体滴度均有显著升高,但无加强应答。显示Vi多糖结合疫苗在诱导小鼠血清IgG应答方面有加强效应。 相似文献
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Sixty-eight Bordetella pertussis isolates (obtained between 1994 and 2004 from the province of Ontario in Canada) were compared by the following phenotypic and genetic analyses: serotyping; pulsed-field gel electrophoresis; and partial DNA sequence analysis of their pertactin, pertussis toxin, and fimbriae genes. Although temporal genetic variations were observed among the isolates, which is consistent with the current view that B. pertussis evolves over time, no specific antigenic or genetic type was detected in 48 isolates collected shortly after the introduction of the acellular pertussis vaccine. Further surveillance with clinical data and isolates collected periodically will be required to ensure that any genetic divergence that could affect vaccine efficacy will not be occurring. 相似文献
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《微生物学免疫学进展》2017,(6)
目的根据"动物法则"验证鼠疫疫苗临床有效性免疫学指标为F1抗体效价和V抗体效价,为疫苗的临床有效性评价提供依据。方法将BALB/c小鼠随机分为6组,每组10只。其中5组用含有不同效价的F1抗体和V抗体的鼠疫疫苗IIa期临床人血清腹腔被动免疫小鼠,另外1组为对照;3 h后各实验组分别以6 MLD鼠疫耶尔森菌标准强毒菌株141株(简称鼠疫强毒菌)攻击,对照组以2MLD鼠疫强毒菌攻击,观察期14 d,详细记录14 d内的动物死亡时间,并对攻击小鼠的存活数据进行Kaplan-Meier生存分析和Spearman相关性分析。结果鼠疫疫苗IIa期临床人血清被动保护小鼠结果显示,对照组小鼠在2 MLD强毒菌攻击下的存活率为0.0%,平均存活时间为6.5 d;实验组临床人血清被动免疫小鼠在6 MLD强毒菌攻击下的存活率分别为10.0%、0.0%、40.0%、40.0%、90.0%,小鼠平均存活时间分别提高到9.1 d、9.9 d、11.2 d、11.4 d、13.9 d。F1抗体效价与小鼠平均存活时间具有显著相关性(P=0.000、r=1.000);V抗体效价与小鼠平均存活时间具有显著相关性(P=0.014、r=0.949)。F1抗体效价与小鼠存活率具有显著相关性(P=0.000、r=1.000);V抗体效价与小鼠存活率具有显著相关性(P=0.014、r=0.949)。结论鼠疫疫苗IIa期临床人血清可对小鼠提供被动保护,且保护力水平与F1抗体效价和V抗体效价具有显著相关性。 相似文献
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Massis LM Braga CJ Sbrogio-Almeida ME Lauand C Newton SM Klebba PE Ferreira LC 《Memórias do Instituto Oswaldo Cruz》2008,103(6):606-610
In the present study we investigated the flagellin-specific serum (IgG) and fecal (IgA) antibody responses elicited in BALB/c mice immunized with isogenic mutant derivatives of the attenuated Salmonella enterica serovar Typhimurium (S. Typhimurium) SL3261 strain expressing phase 1 (FliCi), phase 2 (FljB), or no endogenous flagellin. The data reported here indicate that mice orally immunized with recombinant S. Typhimurium strains do not mount significant systemic or secreted antibody responses to FliCi, FljB or heterologous B-cell epitopes genetically fused to FliCi. These findings are particularly relevant for those interested in the use of flagellins as molecular carriers of heterologous antigens vectored by attenuated S. Typhimurium strains. 相似文献
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The cell mediated immune response (CMI) against pertussis antigens following vaccination with the traditional Danish whole cell pertussis vaccine (WC-P) and the Japanese acellular pertussis vaccine (A-PV) JNIH-3 was studied in four adult human volunteers. Vaccination with the A-PV induced an in vitro proliferative response of peripheral blood lymphocytes to pertussis toxin (PT) subunits S2-S4, S3-S4 and S5 and the filamentous hemagglutinin (FHA), and a better serological response to native PT, detoxified PT (dPT) and FHA than the WC-PV. The induced CMI and serological response were followed over a period of 17 weeks, and were not seen to decline during this period. Further, an in vitro proliferative response to Bordetella pertussis agglutinogen 2 and 3 were demonstrated using lymphocytes from recently and not-so-recently pertussis-vaccinated adults. 相似文献