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460 Local control of ε-gene expression in B cells of the nasal mucosa in hay fever patients following allergen-challenge
Abstracts
J ALLERGY CLIN IMMUNOL VOLUME 97, NUMBER 1, PART 3
457
458
IFN- T T r e a t m e n t R e s u l t s i n t h e F u n c t i o n a l A l t e r a t i o n of A n t i g e n - S p e c l f i c C D 4 ÷ T cells. G Lack MD. A Kupfer MD. A Oshiba MD. E Hamelmann MD. KL Bradley. H Kuvfer MD. J Loader. G Larsen MD. EW Gelfand MD. Denver, CO We investigated the effects of inhaled IFN-T on IgE regulation by CI~* T cells in a murine model of allergic sensitization. BALB/c mice were sensitized to OVA via the airways over 10 days and received nebulized IFN-3' beginning 3 days prior to and during the sensitization period. Two days after the last inhalation, animals were sacrificed and CD4 ÷ T cells were positively selected with irranunomagnetic beads; OVA-specific B cells (OVA + B cells) were isolated using OVA-coated beads and OVA" B cells were isolated from the residual cells using beads coated with anti-kappa antibody. CD4 + T cells (lxl06) from OVA-treated mice were co-cultured with 106 OVA + or OVA" B cells in the presence of IL-4 and the supernates were analyzed 14 days later for polyclonal and anti-OVA IgE by ELISA. OVA + B cells produced high levels of anti-OVA IgE whereas OVA" B cells produced polyclonal IgE. The addition of 106 CIM + T cells from OVA/IFN-3' treated mice significantly inhibited both anti-OVA lgE production by OVA÷ B cells and polyclonal IgE production by OVA" B cells. Addition of anti-IFN-T antibody reversed the CD4-mediated inhibition of IgE production by OVA" B cells but did not reverse the inhibition of anti-OVA IgE production by OVA+ B cells. CD4 + T cells from non-sensitized animals treated with IF'N- 3, alone did not inhibit the production of either polyclonal or anti-OVA lgE, indicating that in vivo allergen exposure is required for CD4"mediated inhibition of IgE production by IFN-y. We conclude that through alterations in CD4+ T cell function, IFN- T treatment inhibits both polyclonal and allergen-specific IgE production, the former being IFN-3' dependent, the latter IFN- T independent. Induction of inhibitory activity by IFN-3' treatment was antigen-dependent.
459
Increased hone marrow progenitor growth after a l l e r g e n c h a l l e n g e in dogs depends on a serum factor MD Inman MD PhD. R Ellis. J Wattle. CG Lane. M D~thlbac/<"PhD. ~A Denbur¢, MD, PM Q'Byrne MB. Asthma Research Group, MeMaster University, Hamilton, ONT, CANADA,and "Astra Draeo, SWEDEN.
460
We have reported that serum taken from dogs at the time of allergen induced airway hyperresponsiveness produces increased growth of granulocyte-macrophage progenitors when added to naive (preallergen) bone marrow. To examine whether this response depended solely on exogenous serum, four dogs who developed (airway responders), and 4 dogs who did not develop allergen-induced airway hyperresponsiveness(controls) were studied in matched pairs. Serum samples taken from each dog prior to and 24h after Ascaris Suum or diluent airway challenge, were added to autologous and matched naive bone marrow cells. Non-adherent mononuclear bone marrow cells were incubated for 8 days in the presence of the serum and recombinant canine GM-CSF, and GM-CFU numbers were counted. Post-allergen challenge serum taken from airway responder dogs produced significant increases (39-42%) in progenitor growth in both aspirates compared to pre-allergen challenge serum (p<0.05). Postallergen challenge serum taken from control dogs did not affect progenitor growth in either aspirates, compared to pre-allergen challenge serum (p>0.05). These findings suggest that allergen induced increases in bone marrow progenitor formation depend solely on a serum hemopoietic factor(s) released post-allergen challenge, and do not depend on a constitutive marrow upregulation in responsiveness to the factor(s).
297
Effect of Acellular Pertussis Vaccine (APV) on the Development of Allergic Sensitization to Environmental A l l e r g e n s , A H A s s a ' a d M D . K Sebastian. M B L i e d MD~ Cincinnati. O H Exposure to pertussis toxin due to infection or vaeeinetion with whole cell pertussis vaccine may increase the serum 1BE,and predispose to sensitization to the prevalent environmental allergens. Since APV also contains Iamussis toxin, it may cause this side effect. As part of,,,, epidemiologic study conducted in the fall, adult hospital employees were randomized to receive either a divalent APV, composed of the pertussis toxin and the filamentous hemagglutinin, or a meningococeal vaccine as a control. Serum specific IgE to cat, dust, Alternaria, and ragweed were measured on sera collected pre-, and one month post- immunization from 51 randomly selected individuals in the APV group, and 49 in the control group. The measurementswere done simultaneouslywith a radioimmunoassayperformed in duplicates. Both groups had no significant change in the serum specific IgE to cat, dust, or ragweed, but showed a significant rise of serum specific lgE to Alternaria one month post immunization. The table shows the results (mean _+SEM) for Alternaria specific lfrE expressed as ~ r , ent of total counts: APV ( n = S t )
Ctmtm~ (n -
49)
Pre
Pest
p
Pr*
Pod
p
0.79 % 4-0.11
0.9 % +_0.11
0.006
0.74% _+0.16
1.02 % _+0.16
0.008
This finding may be due to the seasonal rise in Alternaria specific IgE that accompanies its prevalence during the fall in our regi~m. This study demonstrated that APV given to adults did not predispose to an increase of allergen specific IgE more than a control vaccine.
Local control of c-gene expression in B cells of t h e n a s a l m u c o s a in h a y f e v e r p a t i e n t s f o l l o w i n g a l l e r g e n challenge. SR Durham t. HJ Gould I. CP ThienesI. MR Jacobson I. K MasuyamaI, S Rak2. O Lowhagen2. E Schotman3-and O Humid,3 tLondon UK, 2Gothenburg Sweden, "~Montreal Canada. Topical corticosteroids inhibited allergen induced early and late nasal responses, T cell activation and local interleukin-4 (IL-4) messenger RNA (mRNA) expression in the nasal mucosa (Masuyama K et al, Immunology 1994;82:192). Late responses are known to be lgE-dependent. In 21 patients who underwent a double-blind placebo controlled trial of 6 weeks treatment with fiuticasone propionate aqueous nasal spray (FP) 200meg daily, nasal provocation with grass pollen (Phlegm1pratense 1000Bp.) was performed with biopsy of the inferior turbinate at baseline and 24h after challenge, lgE synthesis was assessed by in situ hybridisation employing riboprobes coding for the IgE heavy chain (Ce) and the I region (IE) 5' to Ce (expressed only in germ-line s transcripts which precede mature Cs mRNA transcription during IgE switch recombination). A control lgG (C7) probe was also used. B cell numbers were assessed using immunostaining with anti-CD20 antibody (APAAP). Cs (p<0.01) but not Is mRNA+ cells were increased at baseline in atopic rhinitics. Following allergen highly significant increases in both lc (p=0.004) and Cs (p=0.006) were observed whereas Cy (p=0.03) mRNA+ cells decreased. These effects were inhibited by FP (all p<0,01). CD20+ B cells did not increase after allergen and were not affected by FP. These results are consistent with the view that allergen-induced IL-4 dependent IgE switch recombination by B cells may occur as a local event in the nasal mucosa which is inhibited by topical corticosteroids.
J ALLERGY CLIN IMMUNOL VOLUME 97, NUMBER 1, PART 3
457
458
IFN- T T r e a t m e n t R e s u l t s i n t h e F u n c t i o n a l A l t e r a t i o n of A n t i g e n - S p e c l f i c C D 4 ÷ T cells. G Lack MD. A Kupfer MD. A Oshiba MD. E Hamelmann MD. KL Bradley. H Kuvfer MD. J Loader. G Larsen MD. EW Gelfand MD. Denver, CO We investigated the effects of inhaled IFN-T on IgE regulation by CI~* T cells in a murine model of allergic sensitization. BALB/c mice were sensitized to OVA via the airways over 10 days and received nebulized IFN-3' beginning 3 days prior to and during the sensitization period. Two days after the last inhalation, animals were sacrificed and CD4 ÷ T cells were positively selected with irranunomagnetic beads; OVA-specific B cells (OVA + B cells) were isolated using OVA-coated beads and OVA" B cells were isolated from the residual cells using beads coated with anti-kappa antibody. CD4 + T cells (lxl06) from OVA-treated mice were co-cultured with 106 OVA + or OVA" B cells in the presence of IL-4 and the supernates were analyzed 14 days later for polyclonal and anti-OVA IgE by ELISA. OVA + B cells produced high levels of anti-OVA IgE whereas OVA" B cells produced polyclonal IgE. The addition of 106 CIM + T cells from OVA/IFN-3' treated mice significantly inhibited both anti-OVA lgE production by OVA÷ B cells and polyclonal IgE production by OVA" B cells. Addition of anti-IFN-T antibody reversed the CD4-mediated inhibition of IgE production by OVA" B cells but did not reverse the inhibition of anti-OVA IgE production by OVA+ B cells. CD4 + T cells from non-sensitized animals treated with IF'N- 3, alone did not inhibit the production of either polyclonal or anti-OVA lgE, indicating that in vivo allergen exposure is required for CD4"mediated inhibition of IgE production by IFN-y. We conclude that through alterations in CD4+ T cell function, IFN- T treatment inhibits both polyclonal and allergen-specific IgE production, the former being IFN-3' dependent, the latter IFN- T independent. Induction of inhibitory activity by IFN-3' treatment was antigen-dependent.
459
Increased hone marrow progenitor growth after a l l e r g e n c h a l l e n g e in dogs depends on a serum factor MD Inman MD PhD. R Ellis. J Wattle. CG Lane. M D~thlbac/<"PhD. ~A Denbur¢, MD, PM Q'Byrne MB. Asthma Research Group, MeMaster University, Hamilton, ONT, CANADA,and "Astra Draeo, SWEDEN.
460
We have reported that serum taken from dogs at the time of allergen induced airway hyperresponsiveness produces increased growth of granulocyte-macrophage progenitors when added to naive (preallergen) bone marrow. To examine whether this response depended solely on exogenous serum, four dogs who developed (airway responders), and 4 dogs who did not develop allergen-induced airway hyperresponsiveness(controls) were studied in matched pairs. Serum samples taken from each dog prior to and 24h after Ascaris Suum or diluent airway challenge, were added to autologous and matched naive bone marrow cells. Non-adherent mononuclear bone marrow cells were incubated for 8 days in the presence of the serum and recombinant canine GM-CSF, and GM-CFU numbers were counted. Post-allergen challenge serum taken from airway responder dogs produced significant increases (39-42%) in progenitor growth in both aspirates compared to pre-allergen challenge serum (p<0.05). Postallergen challenge serum taken from control dogs did not affect progenitor growth in either aspirates, compared to pre-allergen challenge serum (p>0.05). These findings suggest that allergen induced increases in bone marrow progenitor formation depend solely on a serum hemopoietic factor(s) released post-allergen challenge, and do not depend on a constitutive marrow upregulation in responsiveness to the factor(s).
297
Effect of Acellular Pertussis Vaccine (APV) on the Development of Allergic Sensitization to Environmental A l l e r g e n s , A H A s s a ' a d M D . K Sebastian. M B L i e d MD~ Cincinnati. O H Exposure to pertussis toxin due to infection or vaeeinetion with whole cell pertussis vaccine may increase the serum 1BE,and predispose to sensitization to the prevalent environmental allergens. Since APV also contains Iamussis toxin, it may cause this side effect. As part of,,,, epidemiologic study conducted in the fall, adult hospital employees were randomized to receive either a divalent APV, composed of the pertussis toxin and the filamentous hemagglutinin, or a meningococeal vaccine as a control. Serum specific IgE to cat, dust, Alternaria, and ragweed were measured on sera collected pre-, and one month post- immunization from 51 randomly selected individuals in the APV group, and 49 in the control group. The measurementswere done simultaneouslywith a radioimmunoassayperformed in duplicates. Both groups had no significant change in the serum specific IgE to cat, dust, or ragweed, but showed a significant rise of serum specific lgE to Alternaria one month post immunization. The table shows the results (mean _+SEM) for Alternaria specific lfrE expressed as ~ r , ent of total counts: APV ( n = S t )
Ctmtm~ (n -
49)
Pre
Pest
p
Pr*
Pod
p
0.79 % 4-0.11
0.9 % +_0.11
0.006
0.74% _+0.16
1.02 % _+0.16
0.008
This finding may be due to the seasonal rise in Alternaria specific IgE that accompanies its prevalence during the fall in our regi~m. This study demonstrated that APV given to adults did not predispose to an increase of allergen specific IgE more than a control vaccine.
Local control of c-gene expression in B cells of t h e n a s a l m u c o s a in h a y f e v e r p a t i e n t s f o l l o w i n g a l l e r g e n challenge. SR Durham t. HJ Gould I. CP ThienesI. MR Jacobson I. K MasuyamaI, S Rak2. O Lowhagen2. E Schotman3-and O Humid,3 tLondon UK, 2Gothenburg Sweden, "~Montreal Canada. Topical corticosteroids inhibited allergen induced early and late nasal responses, T cell activation and local interleukin-4 (IL-4) messenger RNA (mRNA) expression in the nasal mucosa (Masuyama K et al, Immunology 1994;82:192). Late responses are known to be lgE-dependent. In 21 patients who underwent a double-blind placebo controlled trial of 6 weeks treatment with fiuticasone propionate aqueous nasal spray (FP) 200meg daily, nasal provocation with grass pollen (Phlegm1pratense 1000Bp.) was performed with biopsy of the inferior turbinate at baseline and 24h after challenge, lgE synthesis was assessed by in situ hybridisation employing riboprobes coding for the IgE heavy chain (Ce) and the I region (IE) 5' to Ce (expressed only in germ-line s transcripts which precede mature Cs mRNA transcription during IgE switch recombination). A control lgG (C7) probe was also used. B cell numbers were assessed using immunostaining with anti-CD20 antibody (APAAP). Cs (p<0.01) but not Is mRNA+ cells were increased at baseline in atopic rhinitics. Following allergen highly significant increases in both lc (p=0.004) and Cs (p=0.006) were observed whereas Cy (p=0.03) mRNA+ cells decreased. These effects were inhibited by FP (all p<0,01). CD20+ B cells did not increase after allergen and were not affected by FP. These results are consistent with the view that allergen-induced IL-4 dependent IgE switch recombination by B cells may occur as a local event in the nasal mucosa which is inhibited by topical corticosteroids.