Serum IgM antibody and influenza A infection.

Sucrose density gradient ultracentrifugation followed by haemagglutination inhibition for demonstrating specific influenza IgM was evaluated as a means of confirming recent infection with influenza A viruses. Specific IgM antibodies were found in at least one serum obtained from 83% of patients with proven recent infection with influenza A viruses but in none of the sera from 21 individuals without evidence of infection. Influenza IgM antibodies persisted for up to 112 days after infection. The relative merits of detecting specific IgM and complement fixing antibodies for diagnostic purposes are discussed.     

Treatment of mice with human monoclonal antibody 24h after lethal aerosol challenge with virulent Venezuelan equine encephalitis virus prevents disease but not infection

We recently described a Venezuelan equine encephalitis virus (VEEV)-specific human monoclonal antibody (MAb), F5 nIgG, that recognizes a new neutralization epitope on the VEEV E2 envelope glycoprotein. In this study, we investigated the ability of F5 nIgG given prophylactically or therapeutically to protect mice from subcutaneous or aerosolized VEEV infection. F5 nIgG had potent ability to protect mice from infection by either route when administered 24 h before exposure; however, mice treated 24 h after aerosol exposure developed central nervous system infections but exhibited no clinical signs of disease. Infectious virus, viral antigen and RNA were detected in brains of both treated and untreated mice 2-6 days after aerosol exposure but were cleared from the brains of treated animals by 14-28 days after infection. This fully human MAb could be useful for prophylaxis or immediate therapy for individuals exposed to VEEV accidentally in the laboratory or during a deliberate release.     

Non-depleting anti-CD4 antibody not only prevents onset but resolves sialadenitis in NOD mice

The non-obese diabetic (NOD) mouse spontaneously develops lymphocytic infiltrates in the salivary glands (sialadenitis) and provides an useful rodent model of human Sjogren's syndrome (SS). Non-depleting anti-CD4 antibodies have been shown to ameliorate Type 1 diabetes in NOD mice and also vasculitis in MRL/lpr mice. This study shows that a short course of treatment with the non-depleting anti-CD4 monoclonal antibody, YTS 177, completely prevents salivary infiltration and reverses ongoing pathology in the salivary gland.     

APRIL affects antibody responses and early leukocyte infiltration, but not influenza A viral control

A proliferation inducing ligand (APRIL) is implicated in the regulation of class switch recombination to IgA in T-independent B cell responses. Since B cells play an important role in the immunity to influenza A virus and resistance against the virus is partly controlled by T-independent IgA B cell responses, we studied the role of APRIL during an influenza A infection in vivo. APRIL transgenic, wild-type and APRIL deficient mice were intranasally infected with a non-lethal dose of a mouse adapted strain of influenza A. Compared to wild-type mice, APRIL deficient mice showed a twofold reduction in the amount of macrophages in the lungs and a tendency towards decreased granulocyte influx in the early leukocyte recruitment phase. Although the T cell immune response against influenza was unaffected, APRIL Tg mice showed prolonged influenza-specific IgM production and differential class switching. Unexpectedly, the IgA B cell response was completely T helper cell dependent and also not affected by the absence or presence of APRIL. In addition, viral clearance and recovery from the infection was not influenced by APRIL. Combined these results indicate that APRIL affects specific aspects of the anti-influenza response, but plays a limited role in disease recovery.     

Water-deprivation prevents morphine-, but not LiCl-induced, suppression of sucrose intake.

Intake of a saccharin-conditioned stimulus (CS) can be suppressed following pairing with an aversive agent such as lithium chloride (LiCl) or x-rays (referred to as a conditioned taste aversion or CTA), a highly rewarding sucrose solution (referred to as an anticipatory contrast effect), or a drug of abuse such as morphine or cocaine. Although the suppressive effects of LiCl and sucrose are clear examples of aversive and appetitive conditioning, respectively, it is not certain which properties (aversive or appetitive) mediate the suppressive effects of drugs of abuse. It is known, however, that the suppressive effects of a rewarding sucrose US are attenuated when using a caloric sucrose CS in food deprived rats, while LiCl induced CTAs are much less effected. Standard CTA testing typically is conducted in water-deprived rather than food-deprived rats and, although LiCl is known to suppress intake of a sucrose CS in water-deprived rats, the suppressive effects of drugs of abuse have not been evaluated under these conditions. The present experiment, then, compared the suppressive effects of a standard dose of morphine (15 mg/kg) and a matched dose of LiCl (0.009 M) on intake of a sucrose CS in water-deprived and free-feeding rats. The results showed that both drugs suppressed intake in free-feeding subjects, but only the aversive agent, LiCl, reduced CS intake in the water-deprived rats. This finding dissociates the suppressive effects of morphine and LiCl and, in so doing, aligns the suppressive effects of morphine with those of an appetitive sucrose US.     

Prenatal lipopolysaccharide-exposure prevents allergic sensitization and airway inflammation, but not airway responsiveness in a murine model of experimental asthma

BACKGROUND: Epidemiological evidence underlines the impact of prenatal environmental factors on the development of postnatal allergies. In this regard an inverse correlation between lipopolysaccharide (LPS) exposure and development of childhood allergy has been found. OBJECTIVE: To assess the impact of prenatal LPS exposure on the development of postnatal respiratory allergies in a mouse model of experimental asthma. METHODS: Female BALB/c mice were exposed to LPS before conception and during pregnancy. Several weeks after birth offspring were sensitized to ovalbumin (OVA) followed by aerosol allergen challenges. RESULTS: Prenatal, maternal LPS-exposure enhanced neonatal IFN-gamma, but not IL-4 and IL-2 production. OVA sensitization of prenatally LPS-exposed mice was accompanied by a marked suppression in anti-OVA IgG1 and IgE as well as unchanged IgG2a antibody responses, paralleled by a significant reduction in IL-5 and IL-13 levels following mitogenic stimulation of splenic leucocytes. Assessment of bronchoalveolar lavage fluids following allergen challenges revealed a marked reduction in eosinophils and macrophages in these mice. Surprisingly, development of airway hyper-responsiveness, a hallmark of bronchial asthma, was not affected. CONCLUSION: This study provides first experimental evidence that LPS may already operate in prenatal life in order to modulate the development of allergies in the offspring.     

Mouse Mx2 protein inhibits vesicular stomatitis virus but not influenza virus.

Some but not all known Mx proteins possess intrinsic antiviral activity. The mouse genome contains two related interferon-regulated genes, designated Mx1 and Mx2. Mx1 codes for a nuclear 72-kDa protein which selectively interferes with the multiplication of influenza viruses. The Mx2 gene is crippled by a mutation in commonly used laboratory mouse strains and, hence, the antiviral potential of the Mx2 protein was unknown. We have corrected the frameshift mutation in a cloned Mx2 cDNA by site-directed mutagenesis. Expression of the repaired Mx2 cDNA in Swiss mouse 3T3 cells gave rise to an 80-kDa cytoplasmic protein that cross-reacted with antibodies to other Mx proteins. In contrast to the cases of mouse Mx1 and human Mx proteins, permanent cell lines were extremely unstable with respect to Mx2 expression. Analysis at the single-cell level revealed that mouse Mx2 conferred to the transfected cells a high degree of resistance to vesicular stomatitis virus, but had no inhibitory effect on influenza virus. The antiviral potential of mouse Mx2 is thus similar to that of rat Mx2 protein.     

Serum antibody and ocular responses to murine corneal infection caused by Pseudomonas aeruginosa.

The serum antibody response and differential corneal response to primary and secondary infections by Pseudomonas aeruginosa were investigated in DBA/2J (resistant) and C57BL/6J (susceptible) mice, since they respond differently to intracorneal challenge. Using an enzyme-linked immunosorbent assay, we found that naturally resistant DBA/2J mice mounted a significant immunoglobulin M (IgM) and IgG response to P. aeruginosa within 7 days postinfection of one eye; this was subsequently followed by a drop in the IgM response. Of 31 mice, 30 were able to restore corneal clarity within 3 to 4 weeks. However, when C57BL/6J mice were infected intracorneally, their levels of serum antibody developed more slowly than did those of the DBA/2J mice, and they were unable to restore corneal clarity within 8 to 12 weeks. None of the mice from either test strain mounted a detectable serum IgA response to P. aeruginosa over a 90-day holding period. However, infection of the contralateral, normal cornea of mice of both test strains resulted in a heightened IgG response to P. aeruginosa within 30 days after the secondary infection. Many (50%) of the susceptible C57BL/6J mice recovered or exhibited less severe corneal damage within the 30-day holding period. If the C57BL/6J mice were reinfected 60 days after the primary infection instead of after 30 days, most (89%) of the mice had restored corneal clarity within 3 to 6 days. Passive transfer of immune serum from either recovered DBA/2J or C57BL/6J mice to naive C57BL/6J mice resulted in the restoration of corneal clarity in many of the recipients following infection.     

Naloxone prevents the rapid reacquisition but not acquisition of alcohol seeking

Opioid receptors are involved in reinstatement of alcohol seeking, yet there are no reports of their role in reacquisition of an extinguished alcohol seeking response. Here we investigated the effects of the opioid antagonist naloxone on reacquisition and compared these effects with those on acquisition. Rats were trained, extinguished, then retrained to respond for alcoholic beer. Upon retraining, a second group of rats with no prior experience with the contingency between response and reinforcer was trained under the same conditions. Reacquisition was faster than acquisition. Systemic injection of naloxone (1.25 or 5 mg/kg) reduced reacquisition but had no effect on acquisition. These results suggest that reacquisition and acquisition of alcohol seeking have dissociable neurochemical substrates. (PsycINFO Database Record (c) 2012 APA, all rights reserved).     

Detection of influenza C virus but not influenza D virus in Scottish respiratory samples

BackgroundA newly proposed genus of influenza virus (influenza D) is associated with respiratory disease in pigs and cattle. The novel virus is most closely related to human influenza C virus and can infect ferrets but infection has not been reported in humans.ObjectivesTo ascertain if influenza D virus can be detected retrospectively in patient respiratory samples.Study design3300 human respiratory samples from Edinburgh, Scotland, covering the period 2006–2008, were screened in pools of 10 by RT-PCR using primers capable of detecting both influenza C and D viruses.ResultsInfluenza D was not detected in any sample. Influenza C was present in 6 samples (0.2%), compared with frequencies of 3.3% and 0.9% for influenza A and B viruses from RT-PCR testing of respiratory samples over the same period. Influenza C virus was detected in samples from individuals <2 years or >45 years old, with cases occurring throughout the year. Phylogenetic analysis of nearly complete sequences of all seven segments revealed the presence of multiple, reassortant lineages.ConclusionWe were unable to detect viruses related to influenza D virus in human respiratory samples. Influenza C virus was less prevalent than influenza A and B viruses, was associated with mild disease in the young (<2 years) and old (>45 years) and comprised multiple, reassortant lineages. Inclusion of influenza C virus as part of a diagnostic testing panel for respiratory infections would be of limited additional value.     

Infection enhancement of influenza A NWS virus in primary murine macrophages by anti-hemagglutinin monoclonal antibody.

Antibody-dependent enhancement (ADE) of influenza A NWS virus infection was investigated in primary murine macrophages (M phi) using anti-hemagglutinin(HA) monoclonal antibody (mAB). Contrary to previous reports of abortive influenza virus infection in primary M phi, this study demonstrated that the NWS virus replicated productively in both resident peritoneal M phi and thioglycolate-elicited peritoneal M phi providing cleavage of the HA was achieved by trypsin; 5 micrograms/ml of trypsin was the optimum concentration for the induction of infectivity. Under multiple-cycle growth conditions in the presence of mAB at various concentrations in trypsin-containing media, ADE was demonstrated in both M phi in the presence of subneutralizing concentrations of mAB. Flow cytometric analysis showed that the mechanism of virus entry into M phi could be through HA to specific virus receptors, or HA plus antibody to Fc receptors. These results indicate that ADE of the NWS virus infection actually occurs on Fc receptor-bearing primary murine M phi depending on the concentration of antibody in the presence of the appropriate protease for cleavage of viral HA.     

Graft-facilitating doses of ex vivo activated gammadelta T cells do not cause lethal murine graft-vs.-host disease.

The purpose of this study was to examine the ability of gamma(delta) T cells to cause graft-vs.-host disease (GVHD) after allogeneic bone marrow transplantation (BMT) and to determine whether these cells offered any therapeutic advantages relative to alphabeta T cells. Due to the paucity of naive gamma(delta) T cells in mice and humans, gamma(delta), T cells (obtained from alpha(beta) T cell-deficient murine donors) were ex vivo activated and expanded in interleukin (IL)-2 so as to achieve sufficient cell numbers and to serve as a more clinically feasible strategy. After transplantation into lethally irradiated hosts, donor gamma(delta) T cells were detected in target organs of GVHD such as the spleen and intestines 2 weeks after BMT and constituted the primary T cell subpopulation. Large doses (150 x 10(6)) of activated gamma(delta) T cells, which we have previously shown capable of facilitating engraftment in MHC-disparate recipients, failed to cause fatal GVHD in lethally irradiated recipients of MHC-incompatible donor marrow grafts (C57BL/6 [H-2b]-->B10.BR [H-2k] and C57BL/6 [H-2b]-B6D2F1[H-2b/d]). The absence of GVHD was confirmed by histologic analysis of target organs, splenic B cell reconstitution, and appropriate negative selection in the thymus, that were all comparable to those observed in mice transplanted with T cell-depleted BM only. While early splenic reconstitution was attributable to donor gamma(delta) T cells, analysis of durably engrafted chimeras 2 months posttransplant revealed that the vast majority of donor splenic T cells expressed the alpha(beta) T cell receptor. The results of secondary adoptive transfer assays showed that these cells were tolerant of recipient alloantigens in vivo, demonstrating that gamma(delta) T cells did not prevent the subsequent development of donor anti-host tolerance in BM-derived alpha(beta) T cells. When comparatively evaluated, the minimal number of naive alpha(beta) T cells necessary for donor engraftment caused significantly more fatal GVHD than the corresponding minimal dose of activated gamma(delta) T cells and thus had a superior therapeutic index. These studies indicate that doses of activated gamma(delta) T cells that are able to promote alloengraftment do not cause lethal GVHD in mice transplanted with MHC-incompatible marrow grafts.     

Betamethasone prevents virus-induced airway inflammation but not airway hyperresponsiveness in guinea pigs

In this study the effect of betamethasone was investigated in guinea pigs that demonstrate airway inflammation and airway hyperresponsiveness after a viral respiratory tract infection with parainfluenza-3 (PI3) virus. Guinea pigs were pretreated with saline or betamethasone 8 mg/kg intraperitoneally twice a day for five consecutive days, starting on day 0 and ending on day 4. On day 1, the guinea pigs were inoculated with either control solution (medium) or PI3 virus. On day 5, airway responsiveness was measured. Furthermore, a blood sample was taken, lungs were lavaged, blood leucocytes were counted, and bronchoalveolar lavage (BAL) cells were counted and differentiated. Accordingly, the activity of the bronchoalveolar cells was measured by lucigenin-amplified chemiluminescence. In virus-infected guinea pigs the total bronchoalveolar cell number was increased by 44% compared with medium-treated guinea pigs. This was mainly due to the increase in macrophages (70%, P  < 0.05) and eosinophils (344%, P  < 0.001). The increase in both total and differential (macrophages and eosinophils) cell numbers in virus-infected guinea pigs was completely abolished in animals treated with betamethasone. Moreover, betamethasone prevented the decrease in number of blood leucocytes in virus-infected guinea pigs. In contrast, betamethasone did not prevent the increase in airway responsiveness to both histamine (>200%) and methacholine (>100%) after the virus infection. In conclusion, betamethasone treatment prevents virus-induced airway inflammation but not airway hyperresponsiveness in guinea pigs.     

Serum cytokine levels and antibody response to influenza vaccine in the elderly

Cytokines play critical roles in regulating the antibody response to vaccines. We sought to understand the role of endogenous cytokines in the determination of antibody production in the elderly, a group of subjects known to have a lower response rate to vaccination. We found that in a healthy elderly group, only 52% of whom responded to the influenza vaccine, endogenous levels of interleukin 6 (IL-6), IL-10 and gamma interferon (IFNgamma) did not differ statistically significantly between responders and non-responders (responders: n = 27, IL-6 = 293 +/- 101 pg/ml, IL-10 = 882 +/- 240 pg/ml; nonresponders: n = 26, IL-6 = 223 +/- 71 pg/ml, P = 0.57, IL-10 = 445 +/- 148 pg/ml, mean +/- SE, P = 0.14, respectively, and undetectable IFNgamma). Serum levels of these three cytokines were not changed significantly four weeks after vaccination (P < 0.05 for IL-6 and P < 0.01 for IL-10). In addition, there were also no age-dependent differences in serum IL-6 and IL-10 levels.     

Serum neutralizing antibody and lymphocyte transformation responses after influenza B virus infections

Serum neutralizing antibody and influenza B-specific lymphocyte blast transformation responses were measured in 110 adults and children after an influenza B outbreak. Serum neutralizing antibody and lymphocyte blast transformation responses were seen in 67 to 75% of adults and children recently infected (less than 1 year), but significant lymphocyte blast transformation responses were seen in only 25% of those whose infection was remote (greater than or equal to 3 years). The frequencies of influenza B-induced lymphocyte blast transformation and serum neutralizing antibody responses were similar in the adults and children with similar infection histories.     

IgG but not IgM anti-phospholipid antibody binding is temperature dependent

We examined the effect of temperature on the measurement of enzyme-linked immunosorbent assay (ELISA)-defined human polyclonal antiphospholipid antibody. Both IgG and IgM antibodies were easily demonstrable when sera were incubated on phospholipid-coated ELISA plates at 4–22° C. When incubations were done at 37–45° C IgG antibody binding markedly decreased but IgM antibody binding did not. Warming the phospholipid-coated ELISA plate alone, the serum alone, the buffer alone, or the blocking reagent alone had no effect. When the antigen content of the wells was increased fourfold the effect of warming still occurred. The effect of warmth was reversible and was seen with affinity-purified antibody as well as with whole serum. Phospholipid vesicles in suspension, however, absorbed antibody in a dose-dependent fashion at 4, 22, and 42° C. These results indicate that antibody binding to phospholipid is temperature dependent when phospholipid is adherent to the solid phase. Whether the change in IgG-phospholipid interaction results from a change in antigen or in antibody remains unknown.