Characterisation of the primary local and systemic immune response in gnotobiotic lambs against rotavirus infection

This study characterised the primary immune response in gnotobiotic lambs after infection with a lamb rotavirus (RV). Lambs were infected and killed over a 7 week period together with controls. RV-ELISA and neutralising antibodies were determined in serum, nasal secretions, and intestinal scrapings. RV-antibody secreting cells (ASC) were enumerated in blood. Lymphocyte proliferations were determined in blood and gut-associated lymphoid tissues and cytokine expression was analysed in jejunal Peyer's patches (JPPs) and mesenteric lymph nodes (MLNs). Infected lambs cleared the virus by 8-9 days after infection without showing any clinical signs. The first indication of a specific immune response to RV was an increased expression of IL-4 mRNA in the JPPs in the infected group compared to the control group 3 days after infection. Rotavirus-specific IgA ASC in blood and IgA antibodies in serum and nasal secretions were detected from 7 days after infection followed at 10 days after infection by RV-specific IgG ASC and antibodies. Rotavirus-specific IgA antibodies were not detected in intestinal scrapings in the first 10 days after infection, but were detected by 52 days after infection. No RV-specific neutralising antibodies were seen in the intestine during the course of the experiment.     

Laparoscopic surgery and the systemic immune response

New York Online Access to Health (NOAH) is a Web site that provides accurate, timely, relevant, and unbiased full-text health information in both English and Spanish. A joint project of The City University of New York Office of Library Services, The New York Academy of Medicine Library, the Metropolitan New York Library Council, and The New York Public Library, NOAH brings consumer health information to the public in New York City and around the world via the Internet. NOAH is an example of a successful collaboration among different types of libraries (academic, public, medical society) and voluntary health agencies to use new technologies to reach a very broad public. This paper discusses the involvement of the library partners in terms of the management and funding of the site. Web site construction is described including how the information is gathered and organized. Future plans and funding issues for NOAH are considered in terms of the expected increase in the need for consumer health information. NOAH can be reached at: www.noah.cuny.edu.     

Tumor therapy with an antibody-targeted superantigen generates a dichotomy between local and systemic immune responses

Repeated injections of a fusion protein containing the superantigen staphylococcal enterotoxin A (SEA) combined with a Fab fragment of a tumor-specific antibody is a highly efficient immunotherapy for mice expressing lung melanoma micrometastasis. In the present study, the systemic and local immune responses generated by this therapy were analyzed at a cellular level. Two distinct but coupled immune reactions occurred after repeated therapy. Tumor necrosis factor and macrophage inflammatory protein-1 alpha and -1 beta were immediately synthesized, in the absence of T lymphocytes, at the local tumor site in the lung. This was followed by the induction of VCAM-1 adhesion molecule expression on pulmonary vascular endothelial cells. Concurrently, the early response in the spleen was characterized by the induction of selective T cells producing interleukin (IL)-2. The primed and expanded SEA-reactive V beta 3- and V beta 11-expressing T lymphocytes accumulated to the tumor area only after Fab-SEA therapy and were not present in the lung when SEA, Fab fragment, or recombinant IL-2 was injected. The tumor-infiltrating T cells produced large amounts of interferon-gamma, but no IL-2 or Th2 type of lymphokines were detected at the tumor site in the Fab-SEA-targeted antitumor immune response. These results emphasize the necessity to investigate several sites of antigen presentation to elucidate the effects of immunotherapy.     

Local vs. systemic immune and haemostatic response to hip arthroplasty

Local and systemic immune and haemostatic responses were studied in 10 patients, aged 57-78 years, undergoing elective hip arthroplasty. Cytokines, soluble cytokine receptors, interleukin-1 receptor antagonist, soluble adhesion molecules, antithrombin, fibrin, soluble and fibrin D-dimer were analysed in wound drainage blood and in blood taken from the systemic circulation for up to 24 h post-operatively. Wound drainage blood concentrations of cytokines, interleukin-1 receptor antagonist and soluble cytokine receptors were increased compared with those in the systemic circulation except for the soluble interleukin-6 receptor. In wound drainage blood, soluble tumour necrosis factor receptors (P < 0.05), interleukin-1 receptor antagonist (P < 0.05) and interleukin-6 (P < 0.05-< 0.01) increased during the study period. In blood from the systemic circulation interleukin-6 increased (P < 0.05) while the soluble interleukin-6 receptor decreased (P < 0.05) compared with pre-operative values. Concentrations of soluble adhesion molecules did not change. Wound drainage blood showed marked hypercoagulation. After hip arthroplasty pro-inflammatory cytokines and their inhibitors were mainly confined to the local trauma site. A predominance for inhibitors was noted.     

Disclosure of trauma and immune response to a hepatitis B vaccination program.

This study investigated whether emotional expression of traumatic experiences influenced the immune response to a hepatitis B vaccination program. Forty medical students who tested negative for hepatitis B antibodies were randomly assigned to write about personal traumatic events or control topics during 4 consecutive daily sessions. The day after completion of the writing, participants were given their first hepatitis B vaccination, with booster injections at 1 and 4 months after the writing. Blood was collected before each vaccination and at a 6-month follow-up. Compared with the control group, participants in the emotional expression group showed significantly higher antibody levels against hepatitis B at the 4 and 6-month follow-up periods. Other immune changes evident immediately after writing were significantly lower numbers of circulating T helper lymphocytes and basophils in the treatment group. The finding that a writing intervention influences immune response provides further support for a link between emotional disclosure and health. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Humoral and cellular immune response to elastin in patients with systemic sclerosis

Fifty-six patients (age range, 15-79 yr, average, 37.0+/-18.5 yr), with a clinical and/or radiological diagnosis of acute maxillary sinusitis, were prospectively studied with ultrasound (US) and computed tomography (CT). The imaging finding which supported the diagnosis of acute sinusitis with US was the identification of the hyperechoic posterior antral wall through the hypoechoic inflammation. The findings were compared to CT (3 mm axial sections). The sensitivity of US for maxillary sinus disease was found to be 66.7% and the specificity was 94.9%, which were similar to the plain film ones (65.2 and 96.8%, respectively). The results of the present study suggest US as the method of first choice for acute sinusitis of the maxillary antra, particularly for children and pregnant women.     

DNA vaccination with cytokine fusion constructs biases the immune response to ovalbumin

DNA vaccination may work through direct transfection of antigen presenting cells (APC), or by secretion of the encoded protein by muscle or skin cells for uptake by APC. If cytokines are attached to the antigen, they may influence APC or responding T cells to drive the response toward a Th1 or Th2 direction, and/or potentiate it in an antigen-specific manner. To test this concept, expression vectors were constructed containing the ovalbumin (OVA) gene either alone, or linked to cytokine genes including GM-CSF, IFN-gamma, IL-2, IL-4, IL-12, or a sequence encoding nine amino acids of IL-1 beta. These constructs expressed OVA-cytokine fusion proteins in vitro which retained cytokine bioactivity. C57BL/6 mice were injected intramuscularly with the DNA constructs. Little if any OVA-specific antibody was produced in response to any of the DNA constructs, except for OVA-IL-4. However, lymphocytes from BALB/c mice vaccinated with OVA-IL-12 and OVA-IL-1 beta constructs produced more IFN-gamma and less IL-4 during in vitro restimulation assays than did other groups. All constructs elicited OVA-specific cytotoxic responses which were maintained or even increased over 16 weeks. The OVA-IL-12 and OVA-IL-1 beta peptide constructs elicited the strongest cytotoxic responses at 2 weeks postinjection. Cytotoxic responses were seen in all animals, even those lacking OVA-specific Ab, and were not related to Ab level. These studies indicate that the humoral, cytokine, and cytotoxic responses to DNA vaccination can be effectively altered by certain cytokine fusion constructs.     

Adhesion molecules during immune response to exercise

PURPOSE: The extracellular matrix serves as a structural support for the corneal stroma and mediates signaling events that regulate the intracellular environment of stromal keratocytes. We hypothesize that adhesion and injury mediate signal transduction events causing the phosphorylation of tyrosine residues of specific adhesion proteins and that phosphorylation is required for cellular adhesion and migration. METHODS: For the adhesion experiments; primary rabbit stromal fibroblasts were seeded and phosphorylation of tyrosine residues was followed from 1 min to 24 h. For the injury experiments, confluent primary cultures were rendered quiescent, wounded, and tyrosine phosphorylation was followed from 30 s to 6 h. The antibody (py-20) was used to detect proteins phosphorylated on tyrosine residues. We examined changes in the phosphorylation of focal adhesion kinase (FAK), paxillin and cortactin, using immunoprecipitation and Western blot analysis. RESULTS: In the adhesion experiments, the phosphorylation of a 68-kDa protein was detected after 1 min, and the phosphorylation of a 125-kDa protein was not detected until 15 min. These proteins were identified in re-probed blots as paxillin and FAK. In the injury experiments, FAK phosphorylation was detected within 30 s and remained elevated for 6 h when cells were cultured on fibronectin. Both FAK and paxillin phosphorylation were prominent after injury, but unlike FAK phosphorylation, paxillin phosphorylation decreased over time. Phosphorylation was prominent at the wound margin. After wound closure, it returned to background levels. Tyrosine kinase inhibitors, genistein and herbimycin, decreased the number of adherent cells and altered the rate of cell migration after injury, compared to control (DMSO alone). CONCLUSION: The results indicate that injury and cell-matrix interaction mediate the phosphorylation of specific adhesion proteins and that phosphorylation is required for wound repair.     

Characterisation of immune responses in pancreatic carcinoma patients after mutant p21 ras peptide vaccination

This is a study of immune responses generated by mutant ras peptide vaccination of patients with pancreatic adenocarcinoma. Responding T cells from one patient were cloned and two CD4+ T-lymphocyte clones (TLC) specific for the 12 Val peptide and restricted by HLA-DR6 or DQ2 were obtained. These class II molecules have not previously been found to bind or present mutant ras peptides to T cells. The DR6-restricted TLC showed marked cytotoxicity against autologous target cells pulsed with the 12 Val peptide. Target cells pulsed with the control peptide were not killed. Responding T cells from another patient showed cross-reactivity towards the homologous ras peptides. Investigation by limiting dilution analysis (LDA) revealed different T-cell precursor frequencies for the immunising, mutant ras peptide (1:28000), compared with the normal ras peptide (1:110000).     

An early humoral immune response in peripheral blood following parenteral inactivated influenza vaccination

The enzyme-linked immunospot assay was used to examine the humoral immune response in 15 healthy volunteers immunized with either split or subunit inactivated trivalent influenza vaccine containing A/Beijing/353/89 (H3N2), A/Taiwan/1/86 (H1N1) and B/Yamagata/16/88. The rapidity of the individual B-cell and serum antibody response was examined in lymphocyte and serum samples collected at various time intervals after vaccination. A rapid serological response was detected with increases in antibody titre detected in the majority of volunteers by 7-8 days postvaccination. Influenza-specific plasma cells were detected as early as 4 days postvaccination, higher numbers of IgA and IgG antibody-secreting cells (ASC) were observed which peaked at 7-8 days postvaccination. The number of ASCs then declined, with low numbers of cells detected at 11 days postvaccination. Influenza-specific IgA ASCs were predominantly of the IgA1 subclass. This rapid immune response may have a bearing on future vaccination policies of unimmunized 'at risk groups' in times of high influenza activity.     

Viral myocarditis: balance between viral infection and immune response

Myocarditis is an inflammatory disorder of the heart muscle, and is often underdiagnosed in clinical practice. It presents more dramatically in the young with acute heart failure and more insidiously in adults with chronic dilated cardiomyopathy. The etiology is most often viral in North America, with a wide spectrum of natural history. The majority of patients recover spontaneously, but those with persistent ventricular dysfunction face a 20% one-year mortality. Myocarditis initiates as viral disease, and molecular techniques have confirmed viral persistence. The immune response follows as a two-edged sword-both inadequate and excessive immune responses lead to disease. Finally, the myocyte is the target of the above processes, and expresses molecular, cytokine and vascular changes that lead to dilated cardiomyopathy and heart failure. The gold standard for diagnosis still relies on the overly strict Dallas criteria for evaluating myocardial biopsies. Molecular techniques are playing an increasingly important role in both diagnosis and prognosis. Clinical suspicion is still the key towards an early diagnosis. Treatment must be early and persistent-generally supportive initially, with immunosuppression now playing a secondary role in temporizing those with exuberant immune response. Newer treatments for dilated cardiomyopathy such as amlodipine and carvedilol are equally appropriate for postmyocarditis patients. Future treatment may involve specific biological agents, immune therapy, antiviral strategies and molecular gene therapy.     

Primary infection of dogs with Echinococcus granulosus: systemic and local (Peyer's patches) immune responses

Local and systemic lymphocyte proliferation and antibody production were tested in five dogs 35 days after primary experimental infection with Echinococcus granulosus. A significant cell proliferation was demonstrated by [3H] thymidine incorporation in mesenteric, popliteal and/or Peyer's patches (PPs) cells in response to E. granulosus protoscolex or adult worm antigen in three of five infected dogs, but not in five control animals. In contrast, blood mononuclear cells responded very weakly in only two of the infected dogs to parasite antigens. Elevated levels (compared with preinfection status) of protoscolex- and adult worm antigen-specific serum IgG were detected (ELISA) in four of the five dogs 35 days after infection. Furthermore, slightly elevated levels of parasite-specific IgE and IgA were observed in the sera of three and four in four infected dogs, respectively. Specific serum IgM was not significantly higher 35 days after infection than before infection. Local antibody production was studied in vitro using PPs, mesenteric and popliteal cells isolated from three infected and three uninfected dogs by ELISA using adult worm antigen. In two of three cultures of unstimulated PPs cells of infected dogs, parasite-specific IgG was detectable. Parasite-specific IgA and IgM were detected in one of the unstimulated PPs cell culture derived from an infected dog. Following in vitro stimulation with parasite antigen, PPs cells from two infected dogs showed increased parasite-specific IgG and PPs cells of all three infected dogs produced parasite-specific IgA. PPs cells from uninfected dogs did not produce significant quantities of parasite-specific antibodies and cells from mesenteric and popliteal lymph nodes of infected or uninfected dogs neither produced antibodies whilst in in vitro cultures.     

The immunosuppressive peptide of HIV-1: functional domains and immune response in AIDS patients

OBJECTIVES: To study the biological properties of the immunosuppressive peptide (ISU-peptide) of HIV-1, a 17-mer corresponding to the amino-acid domain 583-599 of the transmembrane glycoprotein gp41 of HIV-1. This peptide exhibits sequence homology to the highly conserved ISU-peptide of type C and D retroviruses. Also, to study the immune response against the corresponding gp41 epitope in AIDS patients. DESIGN: The ISU-peptide and control peptides were synthesized and tested for immunosuppressive activity in different in vitro lymphocyte proliferation assays. Antibody responses were tested using a peptide enzyme-linked immunosorbent assay. A new property of the ISU-peptide, inhibition of HIV-1 replication, was investigated using a cytopathogenicity assay. RESULTS: The ISU-peptide of HIV-1 and the immunosuppressive peptides of type C and type D retroviruses possess similar functional properties. They inhibit mitogen-induced and lymphokine-dependent T-lymphocyte proliferation, they are interspecies-reactive, they must be conjugated to a carrier protein in order to be immunosuppressive, and their N-terminal octamers represent the minimal immunosuppressive domain. HIV-infected individuals develop antibodies against an epitope located at the C-terminal end of the ISU-peptide and the number of responders and antibody titres decrease during progression to AIDS. In addition to its immunosuppressive activity, the ISU-peptide of HIV-1 inhibits the cytopathic effect of HIV-1 on human MT4 cells, suggesting interference with virus replication. CONCLUSIONS: The immunosuppressive property of the ISU-peptide suggests that gp41 might contribute to the development of AIDS. The evolutionary conservation of the immunosuppressive domain and the ability of the corresponding ISU-peptide to inhibit HIV replication suggest that this domain plays an important role in the life cycle of HIV-1.     

Cross-talk between gammadelta T lymphocytes and immune cells in humoral response

The role of gamma delta T cells in immunoregulation is largely unknown. In the current study we noted that gamma delta T cells play a positive role in the humoral response. These positively acting gamma delta T cells are required for the successful adoptive cell transfer of the humoral response, as well as for in vitro generation of plaque-forming cells (PFC). The presented results show that gammadelta T cells cause an increase in interleukin-10 (IL-10) production, which partly elucidates the mechanism of action of these cells. However, experiments with cell culture inserts strongly suggest that direct cell-cell contact between immune and gamma delta H-2-compatible regulatory T cells is critical to the exertion of the positive immunoregulatory function of gamma delta cells. The mechanism of cross-talk between these two cell populations is still not clear but we regard as most likely that the positively acting gamma delta T cells may interact with a complex of heat-shock protein-non-polymorphic MHC (IB) on the surface of T helper type 2 and/or B cells. This could provide, by direct cell-cell contact, the cognate recognition between gamma delta T-cell receptors and heat-shock protein-MHC that leads to positive internal signalling in the immune cells.     

Kinetics of local and systemic immune responses to an oral cholera vaccine given alone or together with acetylcysteine

Disruption of the hydrogen bonding network at the interface of Escherichia coli transaldolase by substitution of R300 to a glutamic acid residue resulted in a monomeric enzyme at basic pH values, with almost no change in the kinetic parameters. The stability of the R300A and R300E mutants towards urea and thermal inactivation is similar to that of the wild-type enzyme. X-ray analysis showed that no structural changes occurred as a consequence of the side chain replacement. This indicates that the quaternary structure is not required for catalytic activity nor does it contribute significantly to the stability of the enzyme. The results are not consistent with a proposed half-of-the-sites reaction mechanism.     

Rosette-forming cells during immune response to Toxoplasma gondii in mice

The rosette-forming cell (RFC) response of mice immunized with varying doses of Toxoplasma gondii was studied by immunocytoadherence (ICA). The specificity of ICA in the present system was tested by passive sensitization with hyperimmune serum in vivo and in vitro. A slight increase in RFC was observed with the latter. Prior treatment of spleen cells from immunized animals with rabbit anti-mouse immunoglobulin resulted in total inhibition of ICA. During the primary and the secondary response after 10 days, the number of RFC rose rapidly to reach the peak on the 3rd day. With secondary immunization 30 days later, the peak shifted to the 2nd day. Mice infected with a lower dose of Toxoplasma had a greater number of RFC during the secondary response after 10 days than with a larger dose.     

Oral immunization with recombinant Norwalk virus-like particles induces a systemic and mucosal immune response in mice

Recombinant Norwalk virus-like particles (rNV VLPs) produced in insect cells were evaluated as an oral immunogen in CD1 and BALB/c mice by monitoring rNV-specific serum total and subclass immunoglobulin G (IgG) and intestinal IgA responses. Dose and kinetics of response were evaluated in the presence and absence of the mucosal adjuvant cholera toxin (CT). rNV-specific serum IgG and intestinal IgA were detected in the absence of CT, and the number of responders was not significantly different from that of mice administered VLPs with CT at most doses. The use of CT was associated with induction of higher levels of IgG in serum; this effect was greater at higher doses of VLPs. IgG in serum was detected in the majority of animals by 9 days postimmunization (dpi), and intestinal IgA responses were detected by 24 dpi. In the absence of CT, IgG2b was the dominant IgG subclass response in both mouse strains. Thus, nonreplicating rNV VLPs are immunogenic when administered orally in the absence of any delivery system or mucosal adjuvant. These studies demonstrate that rNV VLPs are an excellent model to study the oral delivery of antigen, and they are a potential mucosal vaccine for NV infections.     

Relationship between local immune response to Helicobacter pylori and the diversity of disease: investigation of H. pylori-specific IgA in gastric juice

In order to evaluate the relationship between local immune response to Helicobacter pylori and the diversity of disease, 77 asymptomatic subjects who underwent a health examination were studied. Helicobacter pylori-specific IgG in serum and H. pylori-specific IgA in gastric juice were measured by ELISA, and the measured IgA titre was classified into two grades, low or high. Histological classification of gastritis was performed according to the Sydney system. Cytokines in gastric juice were also measured, and the cytotoxin-associated gene A (cagA) status of H. pylori was tested by PCR. Of the 65 subjects who were positive for H. pylori-specific IgG in serum, 38 (58.5%) were classified as H. pylori-specific IgA low titre in gastric juice and 27 (41.5%) had high titres. In the IgG-positive, IgA-low group, the rate of peptic ulcers (especially duodenal ulcers) in endoscopic findings was higher (P < 0.05); the score of activity and the density of H. pylori were higher (P < 0.001 and P < 0.05, respectively); the score of metaplasia was lower (P < 0.05); and the level of interleukin-1 beta was lower (P < 0.05) than in the IgG-positive, IgA-high group. The positive rate of the cagA gene was 84.4% and there was no significant difference between the two groups. There were differences in endoscopic and histological findings between the IgG-positive, IgA-low and the IgG-positive, IgA-high groups. It is suggested that persons infected with H. pylori can be divided into two different states of disease according to local immune response.     

Mechanism of action of BCG-tumor cell vaccines in the generation of systemic tumor immunity. II. Influence of the local inflammatory response on immune reactivity

The intradermal injection of a vaccine composed of 10(7) X-irradiated syngeneic hepatocarcinoma line 10 (L10) cells admixed with 10(8) Mycobacterium bovis strain BCG into inbred Sewall Wright strain 2 guinea pigs induces a local inflammatory reaction and effectively immunizes against a contralateral challenge with viable L10 cells. The relationship between the local inflammatory reaction and the generation of tumor immunity was studied. Immunization against L10 was most effective when both L10 cells and BCG were injected into the same site, less effective when they were injected into separate sites with common lymphatic drainage, and not effective when they were injected at totally separate sites. Enzymatic dissociation of dermal vaccination sites revealed that vaccination with BCG and L10 cells combined induced a significantly greater inflammatory response than did vaccination with BCG alone or L10 cells alone; the inflammatory response was also greater than the combination of these individual responses, suggesting that BCG and L10 cells interacted synergistically in the elicitation of an inflammatory response. Sites receiving a combined vaccine of BCG and L10 cells were infiltrated rapidly by inflammatory cells, and surgical excision of these sites as early as 24 hours after vaccine administration did not affect significantly the development of immune responsiveness. However, vaccination sites induced by the injection of BCG and L10 cells at separate but adjacent sites were slowly infiltrated by inflammatory cells, and surgical removal of these sites within 96 hours of vaccination inhibited later immune responsiveness. Quantitative cellular analysis of these inflammatory reactions showed that inflammation was related to tumor-reactive immunity such that the greater the initial inflammatory process, the greater the resistance to tumor challenge.