Independence of measles-specific humoral and cellular immune responses to vaccination

With a larger, independent cohort and more sophisticated measures, we sought to confirm our work that indicated independence of humoral and cellular immunity following measles vaccination. We recruited an age-stratified random cohort of 764 healthy subjects from all socioeconomic strata, all with medical-record documentation of 2 age-appropriate doses of measles-containing vaccine. We quantified measles-specific neutralizing antibody levels and assayed the interferon-γ (IFN-γ) enzyme-linked immunosorbent spot assay (ELISPOT) response to measles virus. We also measured secreted cytokines from the peripheral blood mononuclear cells (PBMCs) in response to measles virus by performing enzyme-linked immunosorbent assays as secondary measures of cellular immune status. The median antibody level and median IFN-γ ELISPOT response were 844 mIU/mL (interquartile range [IQR]: 418-1,752) and 36 (IQR: 13.00-69.00) spot-forming cells (per 2 × 10(5) PBMCs), respectively. We observed only a very weak and negative correlation (Spearman's r(s) or ρ of -0.090 [95% confidence interval: -0.162--0.018]). We observed a similar lack of quantitatively important correlations between the neutralizing antibody level and any of the secondary measures. Our data confirm the independence of humoral and cellular immune responses after the second dose of measles vaccination. As researchers pursue novel measles vaccine and measles vaccine delivery systems, they must not infer that humoral responses predict cellular responses.     

Characteristics of cellular and humoral immune responses after immunization with different rabies vaccines

Three rabies vaccines were compared: 1. the Fermi type vaccine, a phenol-treated suspension of brain tissue from infected sheep; 2. a virus grown in sheep brain, purified from the contaminating material to 85-90% and inactivated with beta-propiolactone; 3. and the MNIIP-74 strain cultured in Japanese quail embryo cells and inactivated with beta-propiolactone. A single immunization of mice with any of the preparations resulted in about 50% inhibition of splenocyte migration after 2 days; by day 15 the inhibition was 95-98%. By day 45 the migration index returned to the initial level. Increased ability to blast transformation in splenocytes was found by day 15, reached the maximum (160 to 212% of the control level taken for 100%) by day 30, and then began to decrease. The most marked change in blast transformation was brought about by the purified cerebral vaccine, while the less marked one by the tissue culture vaccine. The titre of virus-neutralizing antibodies reached a maximum after 15-30 days, 60 days after immunization it dropped twice. The resistance of mice to intracerebral infection with the rabies virus shortly after immunization might be due to cellular protective factors, while at later intervals it correlated with the level of virus-neutralizing antibodies.     

脂质体流感疫苗制备及体液免疫应答的研究

目的：制备脂质体流感疫苗并对其体液免疫效果和保护效率进行研究.方法：采用薄膜法与冷冻干燥法相结合的方法制得多层脂质体;用制得的脂质体流感疫苗和普通疫苗分别免疫小鼠,用血凝抑制实验（HI）和酶联免疫吸附试验（ELISA）对免后2～13周的小鼠血清的抗体水平进行检测,HI检测其特异性抗体,ELISA检测IgG抗体水平并对小鼠的保护效率进行检测.结果：实验中,脂质体流感疫苗在2μg/mouse组和4μg/mouse两个剂量组中,不同免疫时间内其HI值可分别较非脂质体疫苗组高出2～3和4倍,产生抗体的水平经ELISA实验得出较非脂质体疫苗组高出1倍左右,故可激发更高的体液免疫;保护效力为4/6.结论：脂质体流感疫苗能够刺激机体产生更高的体液免疫和更高的保护效力.     

PIKA as an adjuvant enhances specific humoral and cellular immune responses following the vaccination of mice with HBsAg plus PIKA

An adjuvant is usually used to enhance the immune response induced by vaccines. The choice of adjuvant or immune enhancer determines the effectiveness of the immune response. Currently, aluminium （Alum, a generic term for salts of aluminium） is the only FDA-approved adjuvant. Alum predominantly induces the differentiation of Th2 cells and thus mediates an antibody immune response. Therefore, there is an urgent need for new adjuvants that enhance not only humoral but also cellular immune responses. In the present study, we demonstrates that PIKA （a stabilized dsRNA） as an adjuvant directly induces the activation and the proliferation of both B and NK cells in vitro. Injection of PIKA into mice results in the production of cytokines in vivo. In addition, the study demonstrates that PIKA promotes the maturation of bone marrow-derived dendritic cells （BMDCs） including up-regulation of the co-stimulatory molecules CD80, CD86 and CD40, and the induction of cytokines such as IL-12p70, IL-12p40 and IL-6. Importantly, after immunization of mice with HBsAg plus PIKA, the presence of PIKA enhances the titers of HBsAg-specific IgG and HBsAg-specific IFN-γ production. These results demonstrate that PIKA as an adjuvant can promote both humoral and cellular immune responses. These might have an implication in applying PIKA as an adjuvant to be used in the design and development of both therapeutic and preventive vaccines, and used in the clinical study.     

Induction of humoral and cellular immune responses against hepatitis C virus by vaccination with replicon particles derived from Sindbis-like virus XJ-160

A replication-defective, recombinant Sindbis virus vector was utilized in a novel immunization strategy to induce humoral and cellular responses against hepatitis C virus (HCV). The recombinant vector, pVaXJ-E1E2, expressing the gene for HCV glycoproteins E2 and E1, was constructed by inserting the E1E2 gene into the replicon pVaXJ, a DNA vector derived from Sindbis-like virus XJ-160. The defective replicon particles, XJ-E1E2, were produced by transfecting BHK-21^E+Capsid cells, the packaging cell lines for the vector from XJ-160 virus, with pVaXJ-E1E2. Both glycoproteins, E2 and E1, were stably expressed, as indicated by immunofluorescence assay (IFA) and Western blotting. Mice were vaccinated using a prime-boost strategy with XJ-E1E2 particles combined with Freund’s incomplete adjuvant via intramuscular injection at 0 and 2 weeks. HCV-specific IgG antibody levels and cellular immune responses were evaluated by IFA and IFN-γ ELISPOT, respectively. The results showed that the defective XJ-E1E2 particles in combination with Freund’s incomplete adjuvant induced effective humoral and cellular immune responses against HCV glycoprotein E1 or E2, suggesting that a defective Sindbis particle vaccine is capable of eliciting an effective immune response. These findings have important implications for the development of HCV vaccine candidates.     

Immunization with a synthetic multiepitope antigen induces humoral and cellular immune responses to hepatitis C virus in mice

The immunogenicity of a synthetic multiepitope PCX3 antigen, which contains triple tandem repeats of five conserved epitopes from hepatitis C virus (HCV) polyprotein, was studied in BALB/c mice given three intraperitoneal injections of antigen with Freund's adjuvant. Both a strong antibody response and specific cytotoxic T lymphocytes were induced. The specific anti-PCX3 IgG was able to bind HCV particles from hepatitis C patient sera by incubation overnight. In particular, in transgenic mice with chimeric human livers, anti-PCX3 antibody was able to lower the viral load in two of five mice and to eliminate HCV infection in three of five mice by 2 wk after inoculation with HCV-positive serum from patients. These results indicated that the synthetic multiepitope PCX3 antigen elicits a potent humoral and cellular immune response against HCV.     

Adjuvant effects of orally administered saponins on humoral and cellular immune responses in mice

We have described adjuvant effects of orally administered Quillaja saponins on the immune responses of mice fed inactivated rabies antigen (AG). The in vivo lymphocyte proliferation in mice fed antigen + saponin (AG + SAP) was significantly greater than that in mice fed antigen (AG) alone. Further, the mitogen-induced cell proliferative responses in animals primed with AG + SAP was markedly increased compared with those in the AG group. These changes in clonal expansion were associated with an enhanced helper T cell (Th) and B cell co-operation. The in vivo cell proliferation and in vitro mitogen-induced responses of mice fed AG + SAP correlated with enhanced antibody synthesis. In mice fed saponin alone, there were significant increases in clonal expansion and lymphocyte function. Our present data indicate that the immunocompetence in animals fed AG + SAP was indeed evoked by saponins. Cytotoxic T lymphocyte activity in mice fed SAP or AG + SAP was detected 7 days after booster, in contrast to 21 days in mice fed AG alone. The natural killer cell activity in mice fed SAP alone was greatly enhanced and persisted for an extended period of time.     

Quality and kinetics of the antibody response in mice after three different low-dose influenza virus vaccination strategies

The threat of a new influenza pandemic has led to renewed interest in dose-sparing vaccination strategies such as intradermal immunization and the use of adjuvanted vaccines. In this study we compared the quality and kinetics of the serum antibody response elicited in mice after one or two immunizations with a split influenza A (H3N2) virus, using three different low-dose vaccination strategies. The mice were divided into four groups, receiving either a low-dose vaccine (3 microg hemagglutinin [HA]) intradermally or intramuscularly with or without aluminum adjuvant or the normal human vaccine dose (15 microg HA) intramuscularly. Sera were collected weekly after vaccination and tested in the hemagglutination inhibition, virus neutralization, and enzyme-linked immunosorbent assays. The antibody responses induced after intradermal or intramuscular low-dose vaccinations were similar and lower than those observed after the human vaccine dose. However, low-dose adjuvanted vaccine elicited a serum antibody response comparable to that elicited by the human dose, although the second immunization did not result in any increase in cross-reactive hemagglutination inhibition antibodies, and the peak serum antibody response was observed 1 week later than in the other vaccination groups. Our murine data suggest that the low-dose intradermal route does not show any obvious advantage over the low-dose intramuscular route in inducing a serum antibody response and that none of the low-dose vaccination strategies is as effective as intramuscular vaccination with the normal human dose. However, the low-dose aluminum-adjuvanted vaccine could present a feasible alternative in case of limited vaccine supply.     

Virus-specific major MHC class II-restricted TCR-transgenic mice: effects on humoral and cellular immune responses after viral infection

A transgenic mouse expressing MHC class II-restricted TCR with specificity for a lymphocytic choriomeningitis virus (LCMV) glycoprotein-derived T helper cell epitope was developed to study the role of LCMV-specific CD4+ T cells in virus infection in vivo. The majority of CD4+ T cells in TCR transgenic mice expressed the transgenic receptor, and LCMV glycoprotein-specific TCR transgenic CD4+ T cells efficiently mediated help for the production of LCMV glycoprotein-specific isotype-switched antibodies. In contrast, LCMV glycoprotein-specific TCR transgenic mice exhibited a drastically reduced ability to provide help for the generation of antibody responses specific for the virus-internal nucleoprotein, indicating that intramolecular/intrastructural help is limited to antigens that are accessible to B cells on the viral surface. Antiviral cellular immunity was studied with noncytopathic LCMV and recombinant cytopathic vaccinia virus expressing the LCMV glycoprotein. TCR transgenic mice failed to efficiently control LCMV infection, demonstrating that functional LCMV-specific CD4+ T cells – even if activated and present at extremely high frequencies – cannot directly mediate protective immunity against LCMV. Despite the fact that LCMV-primed CD4+ T cells from TCR transgenic mice as well as from control mice showed low MHC class II-restricted cytotoxic activity in vivo, this did not correlate with protection against LCMV replication in vivo. In contrast, CD4+ T cells from TCR-transgenic mice mediated efficient protection against infection with recombinant vaccinia virus. These results further support the need for different immune effector functions for protective immunity against different viral infections.     

Nature of immune lymph node factors stimulating humoral and cellular immune responses

Institute of Immunology, Ministry of Health of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. D. Ado.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 107, No. 4, pp. 460–462, April, 1989.     

Regulatory effects of osteoprotegerin on cellular and humoral immune responses

The interaction between receptor activator of nuclear factor-kappaB ligand (RANKL) and RANK has been reported to regulate immunity in addition to bone metabolism. The aim of this study was to determine if osteoprotegerin (OPG), an inhibitor of the RANKL-RANK interaction and possibly a new drug against osteoporosis, would adversely affect immunity. OPG was used to treat mice developing different models of cellular and humoral immune responses and also in vitro in T and B cell assays. In mice, OPG does not affect cell-mediated reactions such as contact hypersensitivity to the hapten oxazolone and liver damage, granuloma formation, and infectious load induced by mycobacterial infection. However, OPG increases humoral reactions such as the production of IgM, IgG, and IgE against the T cell dependent antigen keyhole limpet hemocyanin and the production of IgM against the T cell independent antigen Pneumovax. In vitro, OPG modestly co-stimulates T cells but does not affect the proliferation of B cells. OPG has modest immunoregulatory effects that seem to be confined to the humoral response to specific antigens.     

Dietary whole glucan particles do not affect antibody or cell-mediated immune responses to influenza virus vaccination in mice

Influenza virus is a serious health concern. β-glucans derived from plants, bacteria, and fungi have been shown to potentiate immune system responses including those elicited by vaccination. However, in these studies β-glucan was administered as an adjuvant in the vaccine preparation. We hypothesized that addition of a commercially available whole glucan particle supplement to the diet would improve immune response to primary and secondary influenza vaccination in mice. β-glucan was added to pelleted diet and fed to mice at concentrations designed to deliver 0 (control), 1.8 or 90 mg·kg(-1)·day(-1) to each mouse. Influenza vaccine was given intramuscularly in the left hindlimb and primary and secondary responses were assessed. Supplementation with β-glucan was not effective in boosting immune responses to the vaccine, either in the primary or secondary vaccination experiments. Surprisingly, addition of particulate β-glucan to the vaccine itself also failed to elicit a greater antibody response. These observations suggest that this particular form of β-glucan is ineffective in boosting immune response to intramuscular influenza vaccination. Further study is warranted to determine if the use of different mouse models, different vaccine delivery systems, or β-glucans purified from different strains of bacteria, fungi, or plants could improve outcomes using this or similar protocols.     

Anthrax protective antigen administered by DNA vaccination to distinct subcellular locations potentiates humoral and cellular immune responses

Based on the hypothesis that immune outcome can be influenced by the form of antigen administered and its ability to access various antigen‐processing pathways, we targeted the 63 kDa fragment of protective antigen (PA) of Bacillus anthracis to various subcellular locations by DNA chimeras bearing a set of signal sequences. These targeting signals, namely, lysosome‐associated membrane protein 1 (LAMP1), tissue plasminogen activator (TPA) and ubiquitin, encoded various forms of PA viz. lysosomal, secreted and cytosolic, respectively. Examination of IgG subclass distribution arising as a result of DNA vaccination indicated a higher IgG1:IgG2a ratio whenever the groups were immunized with chimeras bearing TPA, LAMP1 signals alone or when combined together. Importantly, high end‐point titers of IgG antibodies were maintained until 24 wk. It was paralleled by high avidity toxin neutralizing antibodies (TNA) and effective cellular adaptive immunity in the systemic compartment. Anti‐PA and TNA titers of ≈10⁵ and ≈10³, respectively, provided protection to ≈90% of vaccinated animals in the group pTPA‐PA63‐LAMP1. A significant correlation was found between survival percentage and post‐challenge anti‐PA titers and TNA titers. Overall, immune kinetics pointed that differential processing through various compartments gave rise to qualitative differences in the immune response generated by various chimeras.     

Lymphocyte blastogenic responses to influenza virus antigens after influenza infection and vaccination in humans.

Virus-specific in vitro cell-mediated immune responses were investigated in 20 normal volunteers who were challenged with liver influenza A/VIC/3/75 (H3N2) virus and in 13 volunteers who were vaccinated with inactivated vaccine containing A/VIC and A/NJ/8/76 (HswN1) antigens. Lymphocyte cultures were established from peripheral blood samples obtained prior to and at various times after infection or vaccination. Blastogenesis was determined by [3H]thymidine incorporation after stimulation of cultures with purified, inactivated, whole influenza viruses. Six days after infection, significantly elevated levels of blastogenesis were observed after in vitro stimulation with A viruses of hemagglutinin and neuraminidase subtypes that were the same as (H3N2) or antigenically distinct from (Heq1Neq1 or HswN1) those of the challenge virus, although maximum stimulation was noted with virus of the same hemagglutinin subtype (H3) as the challenge virus. Similar although more prolonged blastogenic responses were noted in lymphocyte cultures from vaccinees who had serum antibody rises after vaccination. The kinetics of these responses suggest that cell-mediated immunity may play a role in early events after infection and vaccination with influenza virus.     

Kinetics of different specific immunological parameters after rabies vaccination in mice.

A significant protection to an intracerebral challenge of 70 mean lethal doses of a standard live rabies virus strain was obtained in BCG-pretreated mice or in normal mice which had been immunized with a single subcutaneous injection of a beta-propiolactone-inactivated rabies vaccine. Concomitantly, levels of delayed-type hypersensitivity (measured in vivo by the footpad test) and serum-neutralizing activity were evaluated at various times after immunization. All immune criteria were significantly augmented in the BCG-pretreated, rabies-immune mice as compared to normal, rabies-immune mice. However, peak levels of protection, delayed-type hypersensitivity, and serum-neutralizing activity did not occur at the same times. For instance, in the BCG-pretreated, rabies-immune mice, delayed-type hypersensitivity peaked on day 7, protection peaked on day 21, and serum-neutralizing activity peaked on day 60. In BCG-pretreated mice, which did not receive the rabies vaccine, positive delayed-type hypersensitivity, some protection, and serum neutralizing activity were observed 4 to 5 weeks after BCG pretreatment. The possible relationships between specific and nonspecific immunity provoked by rabies virus antigens, tissue culture cell-associated antigens (derived from the bovine fetal kidney cells in which the rabies virus was grown, and BCG are discussed.     

Humoral and cellular responses of mice to infection with a cold-adapted influenza A virus variant.

The serum antibody response and four different cellular immune responses (cytotoxic T cells, delayed-type hypersensitivity T cells, natural killer cells, and cytotoxic macrophage levels) induced in CBA/H mice were measured at different times after intranasal inoculation of a cold-adapted (ca) variant of influenza A virus, influenza virus A/Ann Arbor/6/60-ca, or the parental virus, influenza virus A/Ann Arbor/6/60. At the highest dose of virus inoculated (5 log10 50% tissue culture infective doses), all four cellular responses reached high levels in the lungs of both groups of mice, and serum antibody titers were detected on day 20 after inoculation of either virus. However, whereas extensive replication of the parental virus occurred in the mouse lungs, very limited replication of the ca variant was observed. Macroscopically, infection with the parental virus caused gross lung damage, whereas such damage was almost absent in mice inoculated with the ca variant. Inoculation of 2 to 5 log10 50% tissue culture infective doses of the parental virus induced high cytotoxic T-cell responses, whereas only the highest dose of the ca variant caused a clearly significant cytotoxic T-cell response. As an inoculum of 5 log10 50% tissue culture infective doses of the ca variant caused a substantial primary immune response without appreciable lung damage, the avirulence of the ca variant may be primarily related to its limited ability to replicate productively in mouse lungs.     

Comparison of specificities of humoral and cellular immune responses to haptens

In this study we induced humoral and cellular immunity to three isomers of aminosulfanilic and aminobenzoic acids and compared their respective specificities. These studies were facilitated by using a new method for preferentially inducing a cellular immune response to haptens: the haptens were covalently coupled to mycobacteria and injected into animals in incomplete Freund's adjuvant. Humoral antibody was iduced by coupling the same isomers to bovine serum albumin and injecting them into animals in complete Freund's adjuvant. Both groups of animals were examined for hapten-specific cellular immunity (migration inhibition factor, delayed skin tests) and for antibody response (passive hemolysin test). The results show that although both cell-mediated and humoral responses can discriminate the para, meta and ortho isomeric forms of the haptens used, the antibody response demonstrated much less cross-reactivity. The relevance of this finding to different requirements for B and T cells in antigenic recognition is discussed.