The relationship between respiratory syncytial virus and asthma.

Asthma is a chronic inflammatory disease of the lung that is a leading cause of morbidity and mortality in children worldwide. Most infants who experience wheezing episodes also exhibit evidence of an ongoing respiratory viral infection. Respiratory syncytial virus (RSV) is the leading cause of lower respiratory tract infection in children and is a common cause of wheezing in infants and young children. In the past several decades, a number of studies have demonstrated a relationship between infants with severe RSV infections and the subsequent development of asthma later during childhood. This review provides an overview of data that suggests a severe RSV infection early in childhood is linked to development of asthma later in life. In addition, the current and potential future use of various animal models to gain additional insight into the relationship between RSV and asthma is discussed.

Alcian Blue and Pyronine Y histochemical stains permit assessment of multiple parameters in pulmonary disease models.

Utilization of a combined Alcian Blue and Pyronine Y histochemical method for the assessment of multiple parameters in the respiratory tract of various species is described. Acidic mucins were deep blue (sialylated mucins), red (sulfated mucins), or variably purple (mixture of sialylated/sulfated mucins), and differential mucus production was readily detected in a murine respiratory syncytial virus vaccine model of pulmonary inflammation. Elastic fibers stained red in the walls of pulmonary arteries, connecting airways, alveolar septa, and subpleural interstitium. Mast cells had red to red-purple granular cytoplasmic staining. Nuclei were ubiquitously counterstained pale blue. Representative staining was detected in tissues from multiple species, including inbred mice, rats, ferrets, cats, dogs, sheep, and pigs. The fluorescent property of the stained tissues offers additional modalities with which to analyze tissue sections. This histochemical technique detects multiple critical parameters in routine paraffin sections of lung tissue, reduces the need for repeated serial sectioning and staining, and is cost-effective and simple to perform.

Immunopathology in RSV infection is mediated by a discrete oligoclonal subset of antigen-specific CD4(+) T cells.

Vaccination with the respiratory syncytial virus (RSV) attachment (G) protein results in immune-mediated lung injury after natural RSV infection with pathogenic features characteristic of an exaggerated Th2 response. Here we demonstrate that approximately half of the CD4(+) T cells infiltrating the lungs of G-primed mice utilize a single V beta gene (V beta 14) with remarkably limited CDR3 diversity. Furthermore, elimination of these V beta 14-bearing CD4(+) T cells in vivo abolishes the type 2-like pulmonary injury. These results suggest that a novel subset of CD4(+) T cells may be crucial in the development of pathology during human RSV infection and that genetic or environmental factors prior to or at the time of G antigen exposure may affect the commitment of this discrete antigen-specific T cell subset to Th2 differentiation.

Independent regulation of lymphocytic choriomeningitis virus-specific T cell memory pools: relative stability of CD4 memory under conditions of CD8 memory T cell loss.

Infection of mice with a series of heterologous viruses causes a reduction of memory CD8(+) T cells specific to viruses from earlier infections, but the fate of the virus-specific memory CD4(+) T cell pool following multiple virus infections has been unknown. We have previously reported that the virus-specific CD4(+) Th precursor (Thp) frequency remains stable into long-term immunity following lymphocytic choriomeningitis virus (LCMV) infection. In this study, we questioned whether heterologous virus infections or injection with soluble protein CD4 Ags would impact this stable LCMV-specific CD4(+) Thp memory pool. Limiting dilution analyses for IL-2-producing cells and intracellular cytokine staining for IFN-gamma revealed that the LCMV-specific CD4(+) Thp frequency remains relatively stable following multiple heterologous virus infections or protein Ag immunizations, even under conditions that dramatically reduce the LCMV-specific CD8(+) CTL precursor frequency. These data indicate that the CD4(+) and CD8(+) memory T cell pools are regulated independently and that the loss in CD8(+) T cell memory following heterologous virus infections is not a consequence of a parallel loss in the memory CD4(+) T cell population.

The attachment (G) glycoprotein of respiratory syncytial virus contains a single immunodominant epitope that elicits both Th1 and Th2 CD4+ T cell responses.

BALB/c mice immunized with a vaccinia virus expressing the attachment (G) glycoprotein of respiratory syncytial virus (RSV) develop a virus-specific CD4(+) T cell response that consists of a mixture of Th1 and Th2 CD4(+) T cells following intranasal infection with live RSV. Recent work has shown that both Th1 and Th2 CD4(+) T cells are elicited to a single region comprising aa 183-197 of the G protein. To more precisely define the CD4(+) T cell epitope(s) contained within this region, we created a panel of amino- and carboxyl-terminal truncated as well as single alanine-substituted peptides spanning aa 183-197. These peptides were used to examine the ex vivo cytokine response of memory effector CD4(+) T cells infiltrating the lungs of G-primed RSV-infected mice. Analysis of lung-derived memory effector CD4(+) T cells using intracellular cytokine staining and/or ELISA of effector T cell culture supernatants revealed a single I-E(d)-restricted CD4(+) T cell epitope with a core sequence mapping to aa 185-193. In addition, we examined the T cell repertoire of the RSV G peptide-specific CD4(+) T cells and show that the CD4(+) T cells directed to this single immunodominant G epitope use a restricted range of TCR Vss genes and predominantly express Vss14 TCR.

High frequency of virus-specific interleukin-2-producing CD4(+) T cells and Th1 dominance during lymphocytic choriomeningitis virus infection.

Analysis of C57BL/6 mice acutely infected with lymphocytic choriomeningitis virus (LCMV) by using intracellular cytokine staining revealed a high frequency (2 to 10%) of CD4(+) T cells secreting the Th1-associated cytokines interleukin-2 (IL-2), gamma interferon (IFN-gamma), and tumor necrosis factor alpha, with no concomitant increase in the frequency of CD4(+) T cells secreting the Th2-associated cytokines IL-4, IL-5, and IL-10 following stimulation with viral peptides. In LCMV-infected C57BL/6 CD8(-/-) mice, more than 20% of the CD4(+) T cells secreted IFN-gamma after viral peptide stimulation, whereas less than 1% of the CD4(+) T cells secreted IL-4 under these same conditions. Mice persistently infected with a high dose of LCMV clone 13 also generated a virtually exclusive Th1 response. Thus, LCMV induces a much more profound virus-specific CD4(+) T-cell response than previously recognized, and it is dramatically skewed to a Th1 phenotype.

Attrition of T cell memory: selective loss of LCMV epitope-specific memory CD8 T cells following infections with heterologous viruses.

Using a variety of techniques, including limiting dilution assays (LDA), intracellular IFNgamma assays, and Db-IgG1 MHC dimer staining to measure viral peptide-specific T cell number and function, we show here that heterologous virus infections quantitatively delete and qualitatively alter the memory pool of T cells specific to a previously encountered virus. We also show that a prior history of a virus infection can alter the hierarchy of the immunodominant peptide response to a second virus and that virus infections selectively reactivate memory T cells with distinct specificities to earlier viruses. These results are consistent with a model for the immune system that accommodates memory T cell populations for multiple pathogens over the course of a lifetime.

Protective heterologous antiviral immunity and enhanced immunopathogenesis mediated by memory T cell populations.

A basic principle of immunology is that prior immunity results in complete protection against a homologous agent. In this study, we show that memory T cells specific to unrelated viruses may alter the host's primary immune response to a second virus. Studies with a panel of heterologous viruses, including lymphocytic choriomeningitis (LCMV), Pichinde (PV), vaccinia (VV), and murine cytomegalo (MCMV) viruses showed that prior immunity with one of these viruses in many cases enhanced clearance of a second unrelated virus early in infection. Such protective immunity was common, but it depended on the virus sequence and was not necessarily reciprocal. Cell transfer studies showed that both CD4 and CD8 T cell populations from LCMV-immune mice were required to transfer protective immunity to naive hosts challenged with PV or VV. In the case of LCMV-immune versus naive mice challenged with VV, there was an enhanced early recruitment of memory phenotype interferon (IFN) gamma-secreting CD4(+) and CD8(+) cells into the peritoneal cavity and increased IFN-gamma levels in this initial site of virus replication. Studies with IFN-gamma receptor knockout mice confirmed a role for IFN-gamma in mediating the protective effect by LCMV-immune T cell populations when mice were challenged with VV but not PV. In some virus sequences memory cell populations, although clearing the challenge virus more rapidly, elicited enhanced IFN-gamma-dependent immunopathogenesis in the form of acute fatty necrosis. These results indicate that how a host responds to an infectious agent is a function of its history of previous infections and their influence on the memory T cell pool.

Detection of a high frequency of virus-specific CD4+ T cells during acute infection with lymphocytic choriomeningitis virus.

Lymphocytic choriomeningitis virus (LCMV), like many viruses, induces a profound activation and expansion of CD8+ T cells. In contrast, CD4+ T cells do not increase in total number during the acute infection. We show here that mice infected with LCMV have a low but detectable frequency (<1/300) of CD4+ T cells, as detected by IL-2 production in limiting dilution assays, to each of two class II peptides during the peak of the acute LCMV response and into long-term memory. However, during the peak of the acute CD4+ T cell response, >20% of the CD4+ T cells secreted IFN-gamma after stimulation with PMA and ionomycin, and >10% of the CD4+ T cells secreted IFN-gamma after stimulation with the LCMV peptides. Thus, these new sensitive assays reveal a heretofore unappreciated, yet profound Ag-specific CD4+ T cell response during viral infections.

Stability of virus-specific CD4+ T cell frequencies from acute infection into long term memory.

Mice infected with viruses develop long-lasting high frequency memory CD8+ T cell pools, but much less is known about the CD4+ T cell response. FACS analysis revealed the modulation of several activation markers on CD4+ T cells during an acute infection with lymphocytic choriomeningitis virus (LCMV), consistent with an activated cell phenotype. Examination of virus-specific cytokine production using ELISPOT assays showed a significant increase in the number of IFN-gamma-secreting cells in the spleen during an acute LCMV infection. CD8+ T cells made up the majority of the IFN-gamma-producing cells, but analysis of the cell culture supernatants by ELISA showed that the CD4+ T cells produced more IFN-gamma on a per cell basis. Using limiting dilution assays, we examined the CD4+ T cell precursor (Thp) frequency in C57BL/6 mice infected with LCMV. The virus-specific Thp frequency increased from <1/100,000 in uninfected mice to a peak of approximately 1/600 in purified splenic CD4+ T cell populations by 10 days postinfection with LCMV. After the peak of the response, the Thp frequency decreased only about twofold per CD4+ T cell to approximately 1/1200 and remained stable into long term memory. In contrast to the highly activated CD4+ T cells recovered during the acute LCMV infection, the memory CD4+ T cells were maintained at a lower activation state as judged by cell size and ability to secrete IFN-gamma. Thus, like the CD8+ T cell frequencies, the CD4+ T cell frequencies remain elevated after the acute infection subsides and stay elevated throughout long term immunity.

Alpha beta and gamma delta T-cell networks and their roles in natural resistance to viral infections.

Both alpha beta and gamma delta T-cell populations and natural killer (NK) cells include cytotoxic, interferon (IFN)-gamma-producing lymphocytes that actively respond to viral infections. We show here that all three populations can provide "natural resistance" to viruses very early in infection and describe how the T-cell populations are modulated to provide this function. gamma delta T cells were shown to play a role in controlling vaccinia virus (VV) infections, as VV grew to much higher titers in gamma delta T-cell knockout mice than in normal mice 3-4 days post-infection. Our studies of the alpha beta T-cell responses to viruses revealed an interactive network of T cells that is modulated substantially during systemic infections. There is an induction phase associated with a massive virus-specific CD8 T-cell response, an apoptosis phase during which the T cells become sensitized to activation-induced cell death (AICD), a silencing phase, during which the T-cell number and activation state is reduced, and, finally, a memory phase associated with the very stable preservation of virus-specific memory cytotoxic T-lymphocyte precursors (pCTL). Infection of mice immune to one virus with a heterologous virus leads to a selective expansion of memory CTL cross-reacting between the two viruses, but, after homeostasis is again established, there is a quantitative reduction and qualitative alteration of memory to the first virus. Our results suggest that memory alpha beta T cells cross-reactive between heterologous viruses mediate both immunopathology and protective immunity at early stages of the second virus infection. Thus, memory alpha beta T cells can, like gamma delta T cells and NK cells, provide natural immunity to viral infections.

[Glossopharyngeal neuralgia with syncope]. / Glossopharyngeus neuralgia syncopeval.

Glossopharyngeal neuralgia an uncommon craniofacial pain syndrome. An association with syncope is even less common. The authors give a short case report of 74-year old woman, who has glossopharyngeal neuralgia associated with syncope. During attacks 10 secundum asystolia was recorded in the ECG. The electrophysiologic study of heart was normal, the carotid test resulted 3 secundum asystolia without clinical symptoms. The electroencephalogram and computertomogram of brain were normal. Carbamazepine and demand pacemaker were effective in controlling the symptoms of the patient. Finally, aetiology, pathogenesis and treatment of glossopharyngeal neuralgia are discussed.

The CD45RB-associated epitope defined by monoclonal antibody CZ-1 is an activation and memory marker for mouse CD4 T cells.

Monoclonal antibody CZ-1 defines a novel sialic acid-dependent CD45RB-associated epitope. The CZ-1 antigen is expressed on the subpopulation of CD4 T cells that proliferate in response to IL-2. Because IL-2 responsiveness often denotes T cell activation, we examined the expression of the CZ-1 antigen on CD4 T cells taken from mice at various times during an infection with lymphocytic choriomeningitis virus (LCMV). The blast-sized CD4 T cells at Day 6 postinfection were CZ-1+. Further cell surface phenotyping showed that those blast cells activated at Day 6 postinfection were CD45RBdimPgp-1brightMEL-14-. This contrasts with the CZ-1-CD45RBbrightPgp-1bright/dimMEL-14+ resting cell population prior to infection. To determine if memory CD4 T cells continued to express the CZ-1 epitope long after resolution of the LCMV infection, CD4+CZ-1+ and CD4+CZ-1- populations were purified by cell sorting and placed in an in vitro proliferation assay with LCMV-infected antigen-presenting cells. It was found that the CD4+CZ-1+ population contained virtually all of the virus-specific memory. The CZ-1 epitope is therefore both an activation and a memory marker for murine CD4 T cells.

[Possibilities of early diagnosis of autonomic neuropathy in diabetics]. / A cukorbetegek autonom neuropathiájának koraibb kórismézési lehetösége.

The possibilities of the diagnosis of autonomous neuropathy were investigated by the authors in 71 insulin-treated diabetes mellitus patients with the use of dynamic esophageal scintigraphy and determination of cardiac reflex tests. It has been found that the extent and severity of the lesion can be earlier and more exactly recognized with the help of these two methods. Important information can be obtained by the determination of esophageal transit time about the functional condition of vagal nerves supplying gastrointestinal organs. In patients suffering from autonomous neuropathy the esophageal transit time proved to be significantly prolonged (p less than 0.01). The injury of parasympathetic heart innervation is earlier indicated by the pulmo-cardiac reflexes, while the troubles of symphathic functions are shown more responsively by the sustained handgrip test. Cases with considerable degree of orthostatic hypotension were not observed, its incidence may be probably higher only in advanced forms of autonomous neuropathy. The injury of vegetative nerve fibers develops mostly parallel in diabetes mellitus, but partial functional disorders can be also observed. Detailed neurological examinations were performed in 30 patients, and it has been found, that the lesion of peripheral nerves can be only later observed. Specific complications involving the nervous system are more frequent in cases with longer duration of the disease. In the authors opinion, for investigation and follow-up of larger number of diabetic patients, the dynamic esophageal scintigraphy and determination of pulmo-cardiac reflex and sustained handgrip tests seem to be suitable.

[Hyperacute encephalomyelitis following lyssa vaccination]. / Lyssa vakcinációt követö hiperakut encephalomyelitis.

UNLABELLED: A case of hyperacute disseminated encephalomyelitis following lyssa-vaccination is reported. Clinical symptoms occurred on the tenth day after the vaccination and the patient died three days afterward. Demyelinization and glia proliferation was observed histologically in the spinal cord. No alterations could be seen in the peripheric nerves. CONCLUSIONS: 1. The spinal and cerebral forms of the postvaccinial encephalomyelitis can not be sharply distinguished. 2. In the postvaccinial encephalomyelitis there are also severe infiltration, demyelinization and gliosis in the opticus. 3. Post-vaccinial lyssa encephalomyelitis is a rare, but unavoidable complication of the lassa-vaccination. 4. The State Insurance Company have to pay damages.