Macrophage colony-stimulating factor gene transfer into tumor cells induces macrophage infiltration but not tumor suppression

In order to analyze the effect of a high local concentration of macrophage colony-stimulating factor (M-CSF; CSF-1) on tumor growth, the plasmacytoma cell line J558L was transfected with the human M-CSF gene and injected into syngeneic BALB/c mice. In contrast to the parental tumors, M-CSF transfectants were heavily infiltrated by macrophages as evidenced by immunohistochemistry with antibodies to Mac-1 and Mac-3 and by isolation of the macrophages from the tumor. Nevertheless, tumor growth was only slightly affected by M-CSF and M-CSF-producing cells grew as tumor in all cases. The growth retardation of M-CSF-producing cells varied depending on the experiment and seemed to be due to an indirect effect because the growth rate of the cells in vitro had not changed upon gene transfer. Attempts to activate the tumor-infiltrating macrophages for tumor suppression by systemic application of interferon-γ and/or lipopolysaccharide were not successful. Altogether, our results suggest that M-CSF is a potent chemoattractant for macrophages in vivo but alone is not sufficient to activate these macrophages for tumoricidal activity.     

Lymphocyte subsets in Heymann nephritis

Altered ratios of T lymphocyte subsets have recently been reported in some forms of glomerulonephritis, including membranous glomerulonephritis. Heymann nephritis is a model of membranous glomerulonephritis that can be induced in susceptible strains of rat by a single subcutaneous injection of renal tubular antigen in Freund's complete adjuvant. Monoclonal antibodies were used to identify cytotoxic/suppressor and helper/inducer T cells in the blood, spleen, peripheral lymph nodes, and, where relevant, the lymph node draining the antigen injection site in susceptible and nonsusceptible rat strains before and after immunization with renal tubular antigen. A marked interstrain variation in the proportions of T lymphocyte subsets was found, but this did not segregate strains that are susceptible to Heymann nephritis induction from those that are resistant. Neither the development of Heymann nephritis in susceptible strains or immunization of resistant strains with renal tubular antigen was associated with any specific alteration in the T lymphocyte subpopulations.     

A selective inhibitor of thromboxane biosynthesis enhances immediate and inhibits late cutaneous allergic reactions in man

Thromboxanes may be involved in late cutaneous reactions (LCRs). A new selective inhibitor of thromboxane biosynthesis was studied for its effect on immediate skin test reactions (WFRs) and LCRs in nine adult nonatopic volunteers (open study) and eight adult atopic patients (double-blind study vs. placebo). WFRs and LCRs were elicited by intradermal injections of 25 micrograms of compound 48/80 and 200 IU of rabbit anti-human IgE antibodies, and the patients' receiving injections of 0.32, 1.6, 8, 40, and 200 IU of anti-IgE antibodies. Dazoxiben (100 mg orally three times once every 4 hr) increased the diameter of wheals (p less than 0.005 in volunteers; p less than 0.05 in six patients receiving no additional drug treatment), favored their dispersion (lymphangitis/pseudopodia), and caused systemic reactions (headache, chills) in at least two out of eight patients. Edema formation during LCRs was markedly depressed in 13 out of 16 persons. Flare diameter and erythemata during LCRs were not altered. These data suggest a role of thromboxanes and their natural antagonists (e.g., prostacyclin) in the regulation of allergic inflammatory processes.     

Gene transfer of the interleukin (IL)-2 receptor β chain into an IL-7-dependent pre-B cell line permits IL-2-driven proliferation: Tyrosine phosphorylation of Shc is induced by IL-2 but not IL-7

Expression of the interleukin (IL)-2 receptor β chain in the IL-7-dependent pre-B cell line I × N/2B permitted growth in presence of either IL-2 or IL-7, allowing for a direct comparison of intracellular signaling events. Protein tyrosine phosphorylation was essential for IL-2- and IL-7-induced signal transduction since the tyrosine kinase inhibitor herbimycin A blocked proliferation in response to both factors. Western blot analysis of tyrosine-phosphorylated proteins revealed that both IL-2 and IL-7 stimulation led to enhanced phosphorylation of proteins of 170-, 145, 115- and 99-kDa, as well as induction of phosphorylation of a 96-kDa protein. However, a 55- and a 155-kDa protein were only phosphorylated after IL-2 stimulation. The 55-kDa protein specifically phosphorylated by IL-2 could be identified as p52^shc which has recently been shown to be critically involved in Ras activation. Shc tyrosine phosphorylation as a result of IL-2 stimulation was consistently found in CTLL-2 cells and human T lymphoblasts. Taken together our results indicate that the IL-2- and IL-7-stimulated intracellular pathways are partially different and that Shc is a target of IL2-, but not IL-7-, stimulated tyrosine phosphorylation.     

Effect of Topical Indomethacin on Allergen-Induced Dual Skin Reactions

Twelve asthmatics with dual bronchial and skin reactions after allergen challenge received topical treatment with a 5%-indomethacin cream half an hour before and up to 7h after intradermal allergen and histamine injections. The erythema during the first 20 min of the wheal and flare reaction (WFR) was not affected, neither were the diameters of wheals and flares. 40 to 60 min after injection we observed a marked reduction of the erythema in histamine-and allergen-tested skin areas of 10 Patients. This effect lasted up to the 5th h alter injection of high allergen doses. During the hilly developed late cutaneous reactions (LCR) no effect 0f indomethacin on the erythema was observed, the edema of LCR was only insignificantly reduced. These results suggest that the erythema in LCR between the 1st and 4th is caused, at least in part, by local formation of prostaglandins.     

Allergen tachyphylaxis of guinea pigs in vivo: A prostaglandin E mediated phenomenon?

Ovalbumin (OA) sensitized guinea pigs were repeatedly challenged with 1% OA in saline nebulized ultrasonically at the 0, 10, 20, 60 and 70th min. The intensity of bronchial obstruction was measured by body plethysmography. The first three challenges (0, 10, 20 min) caused strong asthmatic reactions in all animals, the last two (60, 70 min) only mild ones in 10 out of 15 animals. The development of this tachyphylaxis was markedly reduced by pretreatment of the animals with cyclooxygenase inhibitors (indomethacin 10 mg/kg intraperitoneally resp. acetylsalicylic acid 10 mg/kg orally 2h before test). The effect of both inhibitors (i.e. inhibition of tachyphylaxis) was abolished by supplementing prostaglandin E₂ as aerosol simultaneously to the allergen (100–200 ng per inhalation). The results suggest that allergen tachyphylaxis we have observed in vivo might be due to synthesis of cyclooxygenase products, e.g. prostaglandin E.Supported by Deutsche Forschungsgemeinschaft (grant Do 240) in part presented at the 17th workshop on pediatric research [Göttingen 1981, Eur. J. Pediatr. 135: 336 (1981)]     

The effectiveness of invitations from general practitioners in recruiting women to mammographic screening

OBJECTIVE: To evaluate the effectiveness of general practitioner patient lists as a means of recruiting women to mammography screening. DESIGN: This study constitutes the first part of a comparative study of two alternative recruitment strategies involving invitation of women identified from: (i) general practitioner lists; and (ii) the electoral roll. SETTING AND SUBJECTS: The subjects were women aged 50-64 listed as patients of the first three private practices that agreed to collaborate with the South Australian (SA) Breast X-Ray Service to recruit by this method. These practices include five locations encompassing a spread of middle and upper class socioeconomic areas in Adelaide's southern suburbs. INTERVENTIONS: In all, 1505 women who had not already attended the SA Breast X-Ray Service were sent a letter of invitation by their general practitioner to attend the Service for a screening mammogram at a specified date and time. MAIN OUTCOME MEASURES: The primary outcome measures were eligibility status (eligible, not eligible) and attendance status (attended, cancelled appointment, failed to attend without notice). RESULTS: Excluding 34 letters that were returned unopened, 10% of invitees were classified as not eligible, mostly because they had had a recent mammogram elsewhere. Of the remaining invitees, 68.6% attended, 8.4% rang to cancel the appointment and 23% failed to attend without prior notice. CONCLUSIONS: This method of recruitment is viable, and it yields high participation rates close to the "Health for All Australians" target of 70% for mammography screening. Furthermore, the actual attendance rate for this population is expected to increase over time, because some of those initially classified as ineligible, or who cancelled or failed to attend, eventually will attend. The success of this method of recruitment will be measured against the relative cost and effectiveness of the electoral roll alternative, currently under investigation by the SA Breast X-Ray Service.     

Mice deficient in PKC theta demonstrate impaired in vivo T cell activation and protection from T cell-mediated inflammatory diseases

In the present study we have characterized T cell-driven immune function in mice that are genetically deficient in PKC theta. In response to simple immunologic stimulation invoked by in vivo T cell receptor (TCR) cross-linking, these mice showed significantly depressed plasma cytokine levels for IL-2, IL-4, IFNγ, and TNFα compared to wild-type (WT) mice. In parallel, spleen mRNA levels for these cytokines were reduced, and NF-κB activation was also reduced in PKC theta knockouts (KO). Injection of allogeneic cells into the footpad of PKC theta deficient mice provoked a significantly diminished local T cell response compared to WT mice similarly challenged. Unlike comparable cells from wild type mice, CD45RBhi T cells harvested from PKC theta deficient mice failed to induce colitis in the SCID-CD45RB cell transfer model of IBD. In another T cell-dependent model of inflammatory disease, PKC theta deficient animals developed far less severe neurologic signs and reduced spinal cord inflammatory cell infiltrate compared to WT controls in the MOG-induced EAE model. A fundamental role for PKC theta in T cell activation and in the development of T cell-mediated inflammatory diseases is indicated by these results.