Allergic eosinophilic gastroenteritis is characterized by elevated total IgE, specific IgE to food antigens, and eosinophilia of tissue and blood. Because the lymphokines IL-4, IL-5, and gamma-interferon, regulate IgE synthesis, and eosinophilopoiesis in vitro, we examined whether there is an imbalance in their production in allergic eosinophilic gastroenteritis. To explore this hypothesis, three adult patients with allergic eosinophilic gastroenteritis were studied. Flow cytometric studies of peripheral blood mononuclear cells from these patients did not reveal evidence of T cell activation or disturbance of T cell numbers or subsets. T cells were capable of normal mitogenic activation in vitro. IL-4 and IL-5 production were markedly elevated with mitogenic stimulation. Most IL-4 and IL-5 production was by CD4+ T cells. Synthesis of IL-5 by CD4+ T lymphocytes in three patients and CD8+ T lymphocytes in two patients occurred in the absence of mitogen. Mitogen-stimulated GM-CSF and gamma-interferon synthesis by CD4+ T cells was normal. Lymphokine mRNA in total cellular RNA derived from endoscopic biopsies was examined by reverse transcription/polymerase chain reaction. Mucosal biopsies from control subjects and most biopsies from allergic eosinophilic gastroenteritis patients contained less than 10(-8) micrograms IL-5 mRNA/1 microgram total cellular mRNA. gamma-Interferon mRNA was not detected by reverse transcription/polymerase chain reaction in biopsies from patients with allergic eosinophilic gastroenteritis but was present in controls. These lymphokine abnormalities are consistent with the elevated IgE and eosinophilia seen in allergic eosinophilic gastroenteritis and suggest that strategies targeting T lymphocytes may be efficacious in treatment of this disease.

SUMMARYThe expression of cytokine mRNA species was determined in liver biopsies from six normal subjects, 18 patients with PBC and 14 patients with hepatitis B e antigen (HBeAg)-positive CHB using a reverse transcriptase-polymerase chain reaction (RT-PCR) technique. cDNA, obtained by reverse transcription using oligo d(T) primers, was amplified by PCR using primers specific for the coding region of seven different cytokines (IL-1, IL-2, IL-4, IL-5, IL-6, interferon-gamma (IFN-°), tumour necrosis factoralpha (TNF-®)). The abundance of some cytokines (IL-2, IL-4, IL-5 and IFN-°) was also estimated by semiquantitative RT-PCR, using as standards dilutions of synthetic cytokine mRNA transcripts, that could be distinguished electrophoretically from respective native cytokine mRNAs. Hepatic inflammation was assessed by a semiquantitative histologic score and by amplification of mRNA for T cell receptor (TCR)-®. mRNAs for IL-1 and IL-6 were detected in only one control liver. In CHB, mRNAs for IL-1, IL-2, IL-4, IL-5 and IFN-°were detected in 43%, 60%, 80%, 20%, and 54% of biopsies, respectively. mRNA for IFN-°and IL-4, but not IL-1, tended to be associated with severe inflammation. In five biopsies semiquantitative analyses revealed increased levels of mRNA for TCR-® and, when transcripts were detectable, high levels of mRNA for IFN-°and IL-4. In PBC, mRNA for IFN-°was detected in 60% of biopsies, but no mRNAs for IL-1, IL-2, IL-4, IL-5, or IL-6, or for TNF-®, were detected. Semiquantitative analyses revealed that absolute levels of mRNA for IFN-°tended to correlate with the severity of hepatic inflammation. The results suggest that: (i) there may be fundamental differences in the roles that cytokines play in the hepatic inflammatory processes of PBC and CHB; and (ii) while hepatic IFN-°mRNA expression is not specific for PBC, IFN-°may play a prominent role in the immunopathogenesis of PBC.

: Crohn's disease (CD) and ulcerative colitis (UC) are idiopathic inflammatory bowel diseases (IBD) that are characterized by chronic intestinal inflammation and are associated with abnormalities of peripheral and mucosal immune function. The aim of our study was to determine whether CD or UC is characterized by discrete profiles of intestinal lymphokine production. Total cellular RNA was isolated from biopsies of healthy controls and from patients with IBD. Messenger RNA transcript levels in biopsies were determined for interleukin-2 (IL-2), IL-4, IL-5, and interferon-γ (IFN-γ), using a quantitative reverse transcriptase polymerase chain reaction method. Compared with inflamed UC mucosa and controls, CD mucosal lesions contained higher IL-2 and IFN-γ mRNA (p < 0.05), which is consistent with a T-helper cell 1 (Th1)-like pattern. In UC, IL-5 mRNA content was higher in involved areas compared with controls (p < 0.05) and inflamed CD lesions (p < 0.05), suggestive of a Th2 pattern. We conclude that the intestinal mucosa of CD and UC have inflammatory responses characterized by discrete T-helper profiles of lymphokines. This strongly suggests that the immunopathogenesis of these two forms of IBD are different.

Cytokines produced in abnormal amounts or patterns contribute to many immunologically mediated human diseases. We describe a competitive reverse transcription-polymerase chain reaction (RT-PCR) assay to measure interleukin (IL)1-2, IL-4, and interferon-gamma (IFN-gamma) mRNAs within the sample. Internal standard cRNAs and native cytokine mRNAs are reverse transcribed and then amplified by PCR in the same reaction tubes to control for tube-to-tube variability in these reactions. In contrast to systems that use a single multigene internal standard cRNA, this method uses separate internal standard cRNAs for IL-2, IL-4, and IFN-gamma, allowing independent dosing of the internal standards, which reduces the number of tubes processed and the amount of starting mRNA required. Internal standards are produced from cytokine cDNAs by the insertion of short segments of DNA. The same oligonucleotide primers are used to amplify internal standard and native cytokine cDNAs. Each internal standard cDNA and its matching native cytokine cDNA are amplified with equal efficiency. The RT-PCR products of the internal standards and native cytokines are distinguished by size. This technique can detect a twofold difference in mRNA levels. Examples of using this technique to measure cytokine mRNAs in peripheral blood mononuclear cells and in bronchoalveolar lavage cells are given.

: Crohn's disease (CD) and ulcerative colitis (UC) are idiopathic inflammatory bowel diseases (IBD) that are characterized by chronic intestinal inflammation and are associated with abnormalities of peripheral and mucosal immune function. The aim of our study was to determine whether CD or UC is characterized by discrete profiles of intestinal lymphokine production. Total cellular RNA was isolated from biopsies of healthy controls and from patients with IBD. Messenger RNA transcript levels in biopsies were determined for interleukin-2 (IL-2), IL-4, IL-5, and interferon-γ (IFN-γ), using a quantitative reverse transcriptase polymerase chain reaction method. Compared with inflamed UC mucosa and controls, CD mucosal lesions contained higher IL-2 and IFN-γ mRNA (p < 0.05), which is consistent with a T-helper cell 1 (Th1)-like pattern. In UC, IL-5 mRNA content was higher in involved areas compared with controls (p < 0.05) and inflamed CD lesions (p < 0.05), suggestive of a Th2 pattern. We conclude that the intestinal mucosa of CD and UC have inflammatory responses characterized by discrete T-helper profiles of lymphokines. This strongly suggests that the immunopathogenesis of these two forms of IBD are different.

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