BACKGROUNDAlzheimer's disease (AD) is a neurodegenerative disorder characterized by overproduction of beta-amyloid (Abeta), with the subsequent pathologic deposition of Abeta which is important for memory and cognition. Recent studies showed murine models of AD and AD patients inoculated with Abeta(1-42) peptide vaccine had a halted or delayed pathological progression of AD. Unfortunately, the clinical phase IIa trial of Abeta(1-42) peptide vaccine (AN1792) was halted prematurely because of episodes of menigoencephalitis in 18 of the vaccinated patients. The vaccination of BALB/c or Tg2576 transgenic mouse with Abeta(1-15) peptide vaccine is safe and the immune effects are satisfactory. This study further characterizes the specific humoral immune responses in adult rhesus monkeys induced by Abeta(1-15) peptide vaccine.

METHODSFive male adult rhesus monkeys were injected intramuscularly with Abeta(1-15) peptide vaccine at baseline and at weeks 2, 6, 10, 14, 18 and 22. The titers and IgG isotypes of the antibody against Abeta(1-42) in serum was measured by Enzyme-linked Immunosorbent Assay (ELISA). The specificity of the antibody against Abeta(1-42) was determined by Western blot. The Abeta plaques in Tg2576 transgenic mouse brain were stained with the antiserum using immunohistochemistry method.

RESULTSAt the eighth week after the vaccination, antibody against Abeta(1-42) began to develop significantly in serum. The titers of the antibody increased following vaccine boosted and reached 1:3840 at the twenty-fourth week, then decreased after the termination of inoculation. The IgG1 was accounted for the highest level in the antiserum pool. The antibody against Abeta(1-42) showed high specificity. The Abeta plaques in Tg2576 transgenic mouse brain were labeled with the antiserum.

CONCLUSIONAbeta(1-15) vaccine can induce vigorously specific humoral immune responses in adult rhesus monkey.

Amyloid beta-Peptides ; immunology ; Animals ; Antibody Formation ; Antibody Specificity ; Immunoglobulin G ; blood ; Immunoglobulin Isotypes ; blood ; Macaca mulatta ; Male ; Peptide Fragments ; immunology ; Vaccination

PURPOSE: A possible involvement of autoimmune mechanism in the pathogenesis of bronchial asthma has been proposed. Recently, alpha-enolase protein was identified as a major autoantigen recognized by circulating IgG autoantibodies in patients with severe asthma. To evaluate a possible pathogenetic significance of these autoantibodies in severe asthma, isotype (IgG, IgA, IgM, and IgE) and IgG subclass (IgG1, IgG2, IgG3, and IgG4) distributions of autoantibodies to recombinant human alpha-enolase protein were analyzed. PATIENTS AND METHODS: We examined serum samples from 10 patients with severe asthma and 7 patients with mild-to-moderate asthma, and 5 healthy controls by immunoblot analysis. Severe asthma was defined as patients having at least 1 severe asthmatic exacerbation requiring an emergency department visit or admission in the last year despite continuous typical therapies. RESULTS: IgG1 was the predominant IgG subclass antibody response to alpha-enolase protein in patients with severe asthma. IgG1 autoantibody to alpha-enolase protein was detected in 7 of 10 patients with severe asthma (70%), 1 of 7 patients with mild-to-moderate asthma (14.3%), and none of 5 healthy controls (0%) (chi-square test; p < 0.05). IgA, IgM, and IgE autoantibodies to alpha-enolase protein could not be detected in patients with severe asthma. CONCLUSION: IgG1 subclass was the predominant type of autoantibody response to alpha-enolase protein in patients with severe asthma, suggests a possibility of IgG1 autoantibody- mediated complement activation in the pathogenesis of severe asthma.
Adult ; Aged ; Asthma/*enzymology/*immunology ; Autoantibodies/*blood/classification ; Autoantigens ; Case-Control Studies ; Complement Activation ; Female ; Humans ; Immunoglobulin G/blood/classification ; Immunoglobulin Isotypes/blood ; Male ; Middle Aged ; Phosphopyruvate Hydratase/*immunology ; Recombinant Proteins/immunology ; Young Adult

BACKGROUNDTo observe the features of serum specific IgA, IgG, IgM antibodies in the acute phase of hemorrhagic fever renal syndrome (HFRS).

METHODSThe nucleocapsid (NP) protein and glycoproteins (GP) of Hantavirus were expressed by recombinant baculovirus, and used as ELISA antigens to test 61 serial sera of 14 acute phase HFRS patients.

RESULTSSeoul like virus RNA were detected from 11 of 14 patients. An early and strong IgA, IgG and IgM antibody response to recombinant NP (rNP) was observed in almost all HFRS cases. The titers of antibody to rNP was apparently higher than that to Rgp. In the early stage, titer of IgG antibody elevated most drastically among all the three classes of antibodies to rNP, followed by IgM and IgA antibody responses. The elevation trend of IgM and IgA antibodies to rNP stayed nearly at the same level, but the IgA titers to rNP were apparently higher than that of IgM. Among the antibodies to rGP, IgA changed distinctly greater than IgG. The elevation trend of IgM could be found during first week after the onset, and the titers dropped gradually after the second week. IgM antibodies of one case who was viral RNA positive were not detected at early stage, but IgA titers were high. The only severe case of the 14 patients kept the lower IgA, IgG and IgM during the whole acute phase.

CONCLUSIONHFRS patients kept an early and strong humoral response to NP and GPs in acute phase of HFRS.IgA could be used together with IgM to improve the diagnostic accuracy.

Acute Disease ; Adult ; Antibodies, Viral ; blood ; Capsid Proteins ; genetics ; immunology ; Female ; Hantavirus ; genetics ; immunology ; Hemorrhagic Fever with Renal Syndrome ; immunology ; virology ; Humans ; Immunoglobulin Isotypes ; blood ; Male ; Viral Core Proteins ; genetics ; immunology ; Viral Envelope Proteins ; genetics ; immunology

The outer membrane proteins (OMPs) of Brucella (B.) abortus have been extensively studied, but their immunogenicity and protective ability against B. abortus infection are still unclear. In the present study, B. abortus Omp28, a group 3 antigen, was amplified by PCR and cloned into a maltose fusion protein expression system. Recombinant Omp28 (rOmp28) was expressed in Escherichia coli and was then purified. Immunogenicity of rOmp28 was confirmed by Western blot analysis with Brucella-positive mouse serum. Furthermore, humoral- or cell-mediated immune responses measured by the production of IgG1 or IgG2a in rOmp28-immunized mice and the ability of rOmp28 immunization to protect against B. abortus infection were evaluated in a mouse model. In the immunogenicity analysis, the mean titers of IgG1 and IgG2a produced by rOmp28-immunized mice were 20-fold higher than those of PBS-treated mice throughout the entire experimental period. Furthermore, spleen proliferation and bacterial burden in the spleen of rOmp28-immunized mice were approximately 1.5-fold lower than those of PBS-treated mice when challenged with virulent B. abortus. These findings suggest that rOmp28 from B. abortus is a good candidate for manufacturing an effective subunit vaccine against B. abortus infection in animals.
Animals ; Antibodies, Bacterial/blood ; Blotting, Western/veterinary ; Brucella Vaccine/*immunology ; Brucella abortus/*immunology ; Brucellosis, Bovine/*immunology/microbiology/*prevention & control ; Cattle ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel/veterinary ; Enzyme-Linked Immunosorbent Assay/veterinary ; Female ; Immunization/veterinary ; Immunoglobulin G/blood ; Immunoglobulin Isotypes/blood ; Membrane Proteins/genetics/*immunology ; Mice ; Mice, Inbred BALB C ; Models, Animal ; Recombinant Proteins/genetics/immunology ; Vaccines, Subunit/immunology

A few members of coronavirus group I which includes porcine epidemic diarrhea virus (PEDV) use porcine aminopeptidase N (pAPN) as a cellular receptor. Cellular receptors play an important role in virus attachment and entry. However, the low permissiveness of PEDV to APN-expressing porcine cell lines has made it difficult to elucidate the role of pAPN in vitro. The purpose of this study was to prove whether the treatment of soluble pAPN could enhance the antibody production against PEDV in guinea pigs, rabbits and sows. The animals (20 guinea pigs, 8 rabbits and 20 sows) were divided into 4 groups. Group A was injected intramuscularly (IM) with soluble pAPN at one hour before intramuscular infection of PEDV on the same site, group B for IM simultaneous injection of pAPN and PEDV, and group C for IM injection of PEDV only. Group D served as a control of pAPN treatment or PEDV infection. Antibody production against PEDV was compared among groups at regular intervals. The results suggested that pAPN could enhance the antibody production against PEDV in guinea pigs and rabbits which are free of pAPN, however, the effect of pAPN treatment in sows was not clearly elucidated.
Animals ; Antibodies, Viral/*blood ; Antibody Formation ; Antigens, CD13/*administration&dosage ; Cercopithecus aethiops ; Coronavirus/*immunology/physiology ; Coronavirus Infections/immunology/*veterinary ; Enzyme-Linked Immunosorbent Assay/veterinary ; Female ; Guinea Pigs ; Immunoglobulin G/*blood ; Immunoglobulin Isotypes ; Injections, Intramuscular ; Pregnancy ; Rabbits ; Solubility ; Swine ; Swine Diseases/*immunology ; Vero Cells/virology

OBJECTIVETo study the regulating effects of a novel CpG oligodeoxynuleotide and the synergistic effect of chitosan-nanoparticles (CNP) with CpG on immune responses of mice, which were used to develop a novel immunoadjuvant to boost immune response to conventional vaccines.

METHODSA novel CpG ODN containing 11 CpG motifs was synthesized and its bioactivities to stimulate the proliferation of lymphocytes of pig in vitro were detected. Then it was entrapped with CNP prepared in our laboratory by the method of ionic cross linkage, and immunized Kunming mice were co-inoculated with paratyphoid vaccine. The peripheral blood was collected weekly from the tail vein of inoculated mice to detect the contents of IgG, IgA, IgM, and specific antibody against salmonella as well as the levels of interleukin-2 (IL2), IL-4, and IL-6 by SABC-ELISA assay. The numbers of leucocytes, monocytes, granuloytes, and lymphocytes were calculated separately using the routine method. The experimental mice were orally challenged with virulent salmonella 35 days after inoculation.

RESULTSThis CpG ODN could remarkably provoke the proliferation of lymphocytes of pig in vitro in contrast with the control (P < 0.05). Compared with those of the control, immunoglobulins, including IgG, IgA, IgM, and specific antibodies to paratyphoid vaccine, increased significantly in sera from the CpG or CpG-CNP-vaccinated mice (P < 0.05). IL-2, IL-4, and IL-6 increased remarkably in sera from immunized mice (P < 0.05). The leucocytes, monocytes, granuloytes, and lymphocytes of the mice immunized with CpG or CpG-CNP were also increased in number (P < 0.05). After the challenge, these immunity values were elevated in the mice vaccinated with CpG or CpG-CNP. The immunized mice all survived, while the control mice fell ill with evident lesions with diffuse hemorrhage in stomach, small intestine, and peritoneum.

CONCLUSIONSCpG ODN entrapped with CNP is a promising effective immunoadjuvant for vaccination, which promotes humoral and cellular immune responses, enhances immunity and resistance against salmonella by co-administration with paratyphoid vaccine.

Adjuvants, Immunologic ; administration & dosage ; pharmacology ; Animals ; Antibodies, Bacterial ; blood ; Cell Proliferation ; Chitosan ; chemistry ; Drug Therapy, Combination ; Enzyme-Linked Immunosorbent Assay ; Immunoglobulin Isotypes ; blood ; Interleukins ; blood ; Lymphocyte Activation ; drug effects ; Lymphocytes ; cytology ; Mice ; Nanoparticles ; chemistry ; Oligodeoxyribonucleotides ; administration & dosage ; pharmacology ; Paratyphoid Fever ; immunology ; prevention & control ; Salmonella ; physiology ; Salmonella Infections, Animal ; immunology ; prevention & control ; Swine ; immunology ; Typhoid-Paratyphoid Vaccines ; immunology

OBJECTIVETo study the effect of Acanthopanax senticosus injection (ASI) on the activities of human tumor necrosis factor (TNF) and natural killer cell (NKC) in the patients with lung cancer and the underlying mechanism.

METHOD73 cases with lung cancer were randomly divided into two groups, namely, the treatment group (n = 39) and observation group (n = 34); 61 cases with or without other diseases were respectively divided into control A (n = 30) and B (n = 31) groups. The patients in treatment group were injected with ASI for 21 days. The activities of human TNF and NKC and the levels of IgG, IgA and IgM were detected respectively.

RESULTAfter injection with ASI the activity of TNF-alpha in treatment group was comparable with that in the two control groups and was significant lower that that in observation group. The activity of TNF-beta and the levels of IgA, IgG and IgM were significantly higher than those in observation group and two control groups (P < 0.01). The activity of NKC was also remarkably higher than observation and two control groups.

CONCLUSIONASI can regulate the cellular immunity and factor, indicating that ASI can be used as an assistant drug to regulate the function of cellular immunity in the patients with lung cancer.

Adult ; Aged ; Eleutherococcus ; chemistry ; Female ; Humans ; Immunity, Cellular ; drug effects ; Immunoglobulin Isotypes ; blood ; Infusions, Intravenous ; Killer Cells, Natural ; drug effects ; Lung Neoplasms ; blood ; immunology ; pathology ; Lymphotoxin-alpha ; metabolism ; Male ; Middle Aged ; Plants, Medicinal ; chemistry ; Polysaccharides ; administration & dosage ; isolation & purification ; pharmacology ; Tumor Necrosis Factor-alpha ; metabolism

Genes encoding for the premembrane and envelope (prME), envelope (E) and nonstructural protein (NS1) of Japanese encephalitis virus (JEV) were cloned. Each protein was expressed in baculovirus expression system. Of the three proteins expressed in baculovirus system, only prME had hemagglutination activity. The prME (72 and 54 kDa), E (54 kDa) and NS1 (46 kDa) proteins could be detected by Western blotting in the recombinant virus infected cells. Immunogenicity of the recombinant proteins obtained from infected Spodoptera frugiperda (Sf-9) cells was examined in mice. The 3 week-old ICR mice immunized intraperitoneally with three recombinant proteins three times were challenged with a lethal JEV. A survival rate was increased from about 7.7% in unimmunized mice to 92.3% in E + prME and only E groups. The complete protection was shown in prME and live vaccine inoculated groups, respectively. We also measured neutralizing antibody and three immunoglobulin subtypes of IgG1, IgG2a and IgG2b in the sera of mice before and after challenge. Titers of IgG1 antibodies were approximately two to three times higher than that of IgG2b antibodies in all the immunized groups as compared to the control group. However, IgG2a antibody level somewhat increased after challenge, indicating T-helper type 1 (Th1) cell response. The results of this study can provide useful information for developing efficacious subunit vaccine against JEV.
Animals ; Antibodies, Viral/blood ; Baculoviridae/genetics ; Blotting, Western ; Cloning, Molecular ; Encephalitis Virus, Japanese/genetics/*immunology ; Encephalitis, Japanese/*immunology/prevention&control ; Female ; Immunization ; Immunoglobulin Isotypes/blood ; Japanese Encephalitis Vaccines/*immunology/standards ; Mice ; Mice, Inbred ICR ; Microscopy, Fluorescence ; Plasmids ; Recombinant Proteins/genetics/immunology ; Viral Envelope Proteins/genetics/*immunology ; Viral Matrix Proteins/genetics/*immunology ; Viral Nonstructural Proteins/genetics/*immunology