TGF-ß-mediated airway tolerance to allergens induced by peptide-based immunomodulatory mucosal vaccination.

We sought to modulate mucosal immune responses using neonatal vaccination to avert the development of allergic airways disease (AAD). Pulmonary pathology in AAD is driven by T helper (TH)2 cytokines, in particular interleukin (IL)4 and IL13, the expression and actions of which are regulated by the transcription factor STAT6. We developed a peptide homolog of STAT6, STAT6-IP. Neonatal mice given, intranasally, STAT6-IP, in an effort to modulate de novo airways immune responses, developed tolerance following subsequent allergen sensitization, with either ovalbumin or ragweed allergens, as demonstrated by reduced TH2 cytokines and specific immunoglobulin (Ig)E and the significant increases in the latency-associated peptide (LAP)(+) T-regulatory (Treg) cell subset and expression of transforming growth factor (TGF)-ß. This regulatory phenotype was transferrable by CD4(+) T cells or CD11c(+) dendritic cells (DCs) derived from STAT6-IP-vaccinated mice. Anti-TGF-ß treatment during allergen sensitization, however, re-established the pro-inflammatory TH2 response. Thus, neonatal STAT6-IP vaccination induces prospective TGF-ß-dependent tolerance to allergen and constitutes a novel highly effective immunomodulatory allergy prevention strategy.

Interleukin-13-dependent bronchial hyper-responsiveness following isolated upper-airway allergen challenge in a murine model of allergic rhinitis and asthma.

BACKGROUND: We have previously shown that isolated allergic sensitization and challenge of the upper airway results in lower-airway inflammation, which supports the concept of the united airways. OBJECTIVE: This study investigates the hypothesis that isolated upper-airway allergic sensitization is sufficient to induce bronchial hyper-responsiveness (BHR), characteristic of asthma, and that IL-13 is an essential mediator in both the upper and lower airways. METHODS: BALB/c mice were sensitized and challenged by intranasal instillation of allergen ovalbumin (OVA) using our standard protocol. BHR to methacholine was determined and inflammation in nares and lung was assessed. RESULTS: Isolated intranasal application of allergen in awake animals resulted in almost exclusive deposition in the upper airways while in anaesthetized mice there was almost equal distribution in the upper and lower airways. We have demonstrated significant BHR to methacholine challenge in animals receiving OVA only in the upper airway. Also noted was concomitant increase in eosinophilic infiltrates in lung and nares as well as increased granulocytes and IL-13 levels in bronchoalveolar lavage (BAL) fluid. Using a polyclonal anti-IL-13 antibody we have shown inhibition of airways inflammation, both in nares and in lung with significant reduction of granulocytes in BAL from anti-IL-13 treated mice (P<0.0001). Anti-IL-13 treatment also abrogates allergen-induced BHR (P<0.01). CONCLUSION: These data suggest that isolated upper-airway allergen deposition initiates allergic responses along the entire airway. IL-13 mediates both airway inflammation and BHR and may play a role in the communication between the upper and lower airways.

HLA-D region antigens in patients with rheumatoid arthritis.

We studied the distribution of HLA-D region antigens in 2 groups of rheumatoid arthritis (RA) patients: those with mild, nonprogressive disease, and those with severe disease. The results demonstrate that DR4 was significantly increased in both RA patient populations. The frequencies of DR1 and DR4-associated DQw7 alleles, however, were different in these 2 groups of patients. DR1 was significantly increased only in patients with mild RA, and DR4-associated DQw7 was significantly increased only in patients with severe disease. The results of the present study, together with previous data from our laboratory and from other investigators on the incidence of HLA-D region antigens in RA, suggest that both DR and DQ (A and B) genes may be important in conferring susceptibility to RA; DR in the mild forms of the disease, and DQ in severe RA.

Cloning of a short HLA-DQ beta locus-specific cDNA probe: typing for DQw specificities.

A short HLA-DQ beta locus-specific (141 bp) probe was cloned from the full-length pII-beta-1 cDNA. Pst 1-digested genomic DNA from homozygous typing cell lines (HTC) was hybridized with this short DQ beta locus-specific, pDQ beta 141, probe. Restriction fragment length polymorphism (RFLP) patterns generated with this DQ beta locus-specific probe were compared with those obtained with the full-length (627 bp) DQ beta, pII-beta-1, probe. The results demonstrate that the RFLP patterns with the pDQ beta 141 probe were very simple, and no crossreacting DR beta and DX beta bands were observed. DQw1, 2, 3 and 4 specificities could each be identified by a single RFLP.

The responsiveness of human papillomavirus upstream regulatory regions to herpes simplex virus immediate early proteins.

We have tested the responsiveness of the upstream regulatory regions (URR) of human papillomaviruses (HPV) to transactivation by the herpes simplex virus type 1 (HSV-1) immediate early proteins ICP4 (Vmw175) and ICP0 (Vmw110), using recombinant plasmids which contain URRs of HPV 1, 11 or 16 upstream of an SV40-promoted CAT gene. Transfection of the HPV-CAT plasmids into mouse cells expressing a temperature-sensitive ICP4 protein enabled us to demonstrate that ICP4 enhances transcription of the HPV 1 and 16 URRs, but not of the HPV 11 URR. Cotransfection of the HPV CAT plasmids with a plasmid encoding ICP0 indicated that only the URR of HPV 16 responded to this protein, and that the response was host cell dependent.