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PubMed1984

Leukocyte accumulation in sparganosis: demonstration of eosinophil and neutrophil chemotactic factors from the plerocercoid of Spirometra erinacei in vivo and in vitro.

In order to determine whether the plerocercoid of Spirometra erinacei itself has eosinophil and neutrophil chemotactic factors, in vivo and in vitro examinations were carried out. We could observe large numbers of eosinophils and neutrophils which accumulated at the injection site of normal guinea pig skin following intradermal injection of soluble extract of plerocercoids of S. erinacei. At 1 hour after the injection, neutrophils appeared at the site, and the cell number reached its peak at 4 hours. Eosinophils appeared rather later than neutrophils (at 2-4 hours), and the number of cells reached its peak at 8 hours after the injection. Eosinophil and neutrophil chemotactic activities were also confirmed in an in vitro system by using a blind-well chemotaxis chamber with a Millipore filter in dose dependent fashion. An eosinophil chemotactic factor (ECF) with molecular weight of approximately 15,000, and two different neutrophil chemotactic factors, one of about same molecular weight as the ECF and the other of low molecular weight, were demonstrated by gel filtration on Sephadex G-200. Furthermore, it was confirmed that those factors were released from parasite by the detection of intensive eosinophil and neutrophil chemotactic activities in the culture supernatants containing plerocercoids.

Animals↗
PubMed1999

Phase I trial of a recombinant vaccinia virus encoding carcinoembryonic antigen in metastatic adenocarcinoma: comparison of intradermal versus subcutaneous administration.

The principal objectives of this trial were twofold: (a) to examine the safety and relative efficacy of intradermal needle injection versus s.c. jet administration of a carcinoembryonic antigen (CEA)-encoding recombinant vaccinia virus (rV-CEA) over a limited dose range and (b) to evaluate CEA-specific immune responses or antitumor effects induced by rV-CEA vaccination. Patients were randomly assigned to one of two groups, depending upon the technique of vaccine administration. All 20 patients received two doses of 10(7) or 10(8) pfu of rV-CEA at a 4-week interval. Toxicity was limited to modest local inflammation at the inoculation site as well as low-grade fever and increased fatigue, each affecting fewer than 20% of the patients. No evidence of CEA-specific lymphoproliferation, interleukin 2 release, delayed-type hypersensitivity, or antibody response was observed. Thus, the efficacy comparison between the two administration techniques was based upon the induction of immune responses to the vaccinia virus vector. Both techniques induced vaccinia-specific lymphoproliferation, interleukin 2 release, and antibody responses of comparable magnitude and frequency as well as protected 80% of patients against pustule formation following vaccinia scarification. Thus, there is no compelling reason to recommend one administration technique over the other based upon toxicity or efficacy. We have selected s.c. jet injection for subsequent trials of rV-CEA based on the ability to accommodate larger injection volumes, enhanced standardization between clinicians, and avoidance of needles that could transmit the vaccine or blood-borne pathogens to health care workers. We recommend use of 10(8) pfu doses for subsequent trials of recombinant vaccinia virus vaccines based upon the favorable toxicity profile and more consistent local pustule formation indicative of an adequate inoculation of live virus. No objective clinical responses to the rV-CEA vaccine were observed among this population of patients with widely metastatic adenocarcinoma.

Adenocarcinoma↗
PubMed1999

Genetically fluorescent melanoma bone and organ metastasis models.

We report here the establishment and metastatic properties of bright, highly stable, green fluorescent protein (GFP) expression transductants of the B16 mouse malignant melanoma cell line and the LOX human melanoma line. The highly fluorescent malignant melanoma cell lines allowed the visualization of skeletal and multiorgan metastases after i.v. injection of B16 cells in C57BL/6 mice and intradermal injection of LOX cells in nude mice. The melanoma cell lines were transduced with the pLEIN expression retroviral vector containing the GFP and neomycin resistance genes. Stable B16F0 and LOX clones expressing high levels of GFP were selected stepwise in vitro in levels of G418 of up to 800 microg/ml. Extensive bone and bone marrow metastases of B16F0 were visualized by GFP expression when the animals were sacrificed 3 weeks after cell implantation. Metastases for both cell lines were visualized in many organs, including the brain, lung, pleural membrane, liver, kidney, adrenal gland, lymph nodes, skeleton, muscle, and skin by GFP fluorescence. This is the first observation of experimental skeletal metastases of melanoma, which was made possible by GFP expression. These models should facilitate future studies of the mechanism and therapy of bone and multiorgan metastasis of melanoma.

Animals↗
PubMed1977

Migration-inhibition response of peripheral leukocytes to tuberculin in cats sensitized with viable Mycobacterium bovis (BCG).

Delayed hypersensitivity reactions to antigens injected intradermally are reported as not occurring in the cat. Cats infected with viable Mycobacterium bovis (BCG) organisms developed a transient migration-inhibition response of their leukocytes to tuberculin. The migration-inhibition response subsided in 6 weeks in spite of the persistance of viable BCG. Significant intradermal reactions to tuberculin 60 days after BCG injection did not occur. The response of the cat differs from that of many mammalian species in which strong in vivo and in vitro delayed hypersensitivity type reactions persist with mycobacterial infections. Despite the lack of continued measurable delayed hypersensitivity, the cat appears to have adequate resistance to mycobacterial infections.

Animals↗
PubMed1994

Anti-tumour effects of interleukin 1 beta: in vivo induction of immunity to B16 melanoma, a non-immunogenic tumour.

The anti-tumour properties of interleukin 1 beta (IL-1 beta) were examined using an intradermal B16 murine melanoma surgical model. B16 cells were injected intradermally on the right ventral side and surgery was performed on days 10-20 to remove the primary tumours. IL-1 beta or vehicle was administered prior to surgery for 5-7 consecutive days. In mice which received only injections of vehicle, survival ranged between 0 and 30% when measured on day 120 after implantation of B16 cells. Mice died of metastases and growth of B16 cells in the thoracic lymph nodes. When mice without metastases were rechallenged with viable B16 cells, only one out of 22 mice (5%) failed to develop tumours. No significant immunity to B16 cells was detected in this group of mice. In contrast, in mice which received injections of IL-1 beta, survival ranged between 70-100% on day 120 after implantation of B16 cells. When IL-1 beta treated mice were rechallenged with viable B16 cells on day 120, 20 out of 32 (63%) mice failed to develop B16 tumours suggesting that some of these mice had immunity to B16 melanoma cells. Moreover, mice with immunity to B16 cells did develop tumours when injected with another syngeneic tumour, MCA 105. In vitro specific immune responses were also demonstrated in spleen cells and sera from mice treated with IL-1 beta, but not in the spleen cells or sera of mice that received only vehicle.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1976

Rat IgE production. II. Primary and booster reaginic antibody responses following intradermal or oral immunization.

Primary and booster IgE antibody responses have been elicited in Hooded Lister rats by the intradermal injection or oral administration of very small quantities of egg albumin. Oral immunization was effected by giving antigen by stomach tube or in the drinking water. The minimum primary dose of antigen found to be effective was 1 mug intradermally and 10 mug orally, administered together with an intraperitoneal injection of B. pertussis adjuvant. In rats immunized with these doses secondary responses could be evoked by giving even smaller quantities of antigen, thus 1 ng intradermally or 1 mug orally without adjuvant. Smaller challenge doses were not tried. Large primary doses of antigen (greater than 100 mug) presented by these routes were, on the other hand, found to be inhibitory to the production of secondary IgE responses, this effect being similar to that observed in previously reported intraperitoneal immunization experiments. By contrast with previous experiments, however, tertiary responses could be obtained following immunization by these routes, and we believe this to be reflection of the absorption of smaller and therefore less inhibitory quantities of antigen. Our results are discussed in relation to the control of IgE antibody production, current concepts of the control of antigen absorption through mucosal barriers, and possible implications of the genesis of naturally occurring IgE responses in man.

Adjuvants, Immunologic↗
PubMed1985

Ultra-long-duration local anesthesia produced by injection of lecithin-coated methoxyflurane microdroplets.

This study was designed to evaluate a new drug delivery system. The authors undertook to determine if microdroplets prepared by encapsulating volatile anesthetics with a membrane of lecithin could be used for local anesthesia. Local anesthesia was determined by monitoring the response of the rat to tail clamping and electrical stimulation of the skin following the intradermal injection of the microdroplets. Microdroplets were prepared from isoflurane, enflurane, halothane, methoxyflurane, diethyl ether, chloroform, and heptane. Although all microdroplet preparations produced local anesthesia, only methoxyflurane microdroplets produced an ultra-long duration of local anesthesia (approximately 24 h). Further characterization of the methoxyflurane microdroplets revealed two important differences from conventional local anesthetics. First, the local anesthetic effect of methoxyflurane reached a plateau that did not change significantly for 20 h while the injection of lidocaine and bupivacaine resulted in a peak effect that returned to baseline within 1 and 3 h, respectively. Second, the anesthetic effect of methoxyflurane remained essentially localized to the site of injection, while the anesthetic effect of lidocaine and bupivacaine migrated 15 cm in less than 1 h. The toxicity and safety of methoxyflurane were evaluated. When administered over the dosage range 1-16% (v/v) intradermally, or by injections into muscle, or by repeat injections every 4 days for 16 days, all animals regained their pretreatment response to painful stimulations, and there was no evidence of gross injury to tissue. Deliberate intravenous injection of 0.8 ml of 6.7% (v/v) methoxyflurane microdroplets had no apparent anesthetic or toxic effect. The present study demonstrates that methoxyflurane microdroplets produce an anesthetic effect that is highly localized, stable in intensity, ultra-long in duration, and reversible.

Anesthesia, Local↗
PubMed2006

Role of the Yersinia pestis plasminogen activator in the incidence of distinct septicemic and bubonic forms of flea-borne plague.

Yersinia pestis is transmitted by fleas and causes bubonic plague, characterized by severe local lymphadenitis that progresses rapidly to systemic infection and life-threatening septicemia. Here, we show that although flea-borne transmission usually leads to bubonic plague in mice, it can also lead to primary septicemic plague. However, intradermal injection of Y. pestis, commonly used to mimic transmission by fleabite, leads only to bubonic plague. A Y. pestis strain lacking the plasmid-encoded cell-surface plasminogen activator, which is avirulent by intradermal or s.c. injection, was able to cause fatal primary septicemic plague at low incidence, but not bubonic plague, when transmitted by fleas. The results clarify a long-standing uncertainty about the etiology of primary septicemic plague and support an evolutionary scenario in which plague first emerged as a flea-borne septicemic disease of limited transmissibility. Subsequent acquisition of the plasminogen activator gene by horizontal transfer enabled the bubonic form of disease and increased the potential for epidemic spread.

Animals↗
PubMed2001

Influence of type 2 diabetes on the inflammatory response in rats.

OBJECTIVES AND DESIGN: To verify whether the inflammatory responses in animals with type 2 diabetes are altered to an extent similar to that in type 1 diabetes. MATERIALS: Male newborn (2 days old) Wistar rats were made diabetic by streptozotocin (160 mg/kg, i.p.) and used 8-10 weeks later (10 rats/group). METHODS: The inflammatory responses were evaluated using paw edema (induced by local injection of carrageenan or dextran), pleurisy (by pleural injection of carrageenan), increases in vascular permeability (induced by intradermal injection of histamine, serotonin and bradykinin) and leukocyte counts in peripheral blood and pleural exudate. RESULTS: Diabetic animals showed reduced inflammatory responses to carrageenan but not to dextran. The increase in vascular permeability induced by serotonin and bradykinin was reduced whereas that to histamine was not altered in diabetic compared to control rats. Although the pleural exudate was reduced, leukocyte counts were similar in diabetic and control rats. Insulin (2 IU, 4 h before), though effective in reducing blood sugar levels, did not restore the altered responses in diabetic rats. In contrast to that in rats with type 1 diabetes, in rats with type 2 diabetes, removal of the adrenal glands restored the reduced inflammatory responses. CONCLUSIONS: Insulin resistance in type 2 diabetic rats led to reduced inflammatory responses, which were partially corrected by adrenalectomy.

Adrenalectomy↗
PubMed1985

Thymosin alpha 1-induced modulation of cellular responses and functional T-cell subsets in mice with experimental autoimmune thyroiditis.

The effects of Ta-1, a peptide constituent of thymosin fraction 5, were studied on murine autoimmune thyroiditis using two congenic strains of mice, B10.Br (Br) and B10.D2 (D2), which are sensitive and resistant to experimental autoimmune thyroiditis (EAT) induction, respectively. EAT was induced by either 2 weekly iv injections of mouse thyroglobulin with adjuvant lipopolysaccharide (LPS) or intradermal injection of thyroglobulin mixed with complete Freund's adjuvant (CFA). The criteria for induction and intensity of thyroiditis were the level of lymphoid infiltration in the thyroid gland and the titer of anti-thyroglobulin antibodies. Ta-1 was given in 5 or 10 daily sc injections in doses ranging from 0.0001 to 0.1 microgram/injection. The injections were commenced at varying intervals from the 1st to the 4th week after immunization. T-Cell subsets in the spleens were determined 2 weeks after the first antigen injection and thyroid infiltration was determined 3 weeks later. Treatment with Ta-1 between the two antigen injections increased the level of thyroiditis in resistant mice, but had no effect in sensitive mice. Treatment for the first 2 weeks had similar effects in resistant mice, but also suppressed thyroiditis in the sensitive strain. Later treatments, during the 3rd and 4th weeks after immunization also revealed immunomodulating properties of Ta-1, with a suppressing effect on thyroiditis in sensitive mice and an enhancing effect in the resistant strain. Both effects of Ta-1 were dose dependent. The effects of Ta-1 on the individual phenotypes were also dose dependent. The dose of 0.01 microgram greatly lowered the percentages of Lyt-2+3+ cells in D2 mice and mildly increased the percentages in Br mice, but did not change the Lyt-1+ cell level in either strain. On the other hand, the dose of 0.001 microgram greatly increased the percentage of Lyt-1+ cells in D2 mice and mildly decreased it in the Br strain, but did not alter the Lyt-2+3+ cell subset in either strain. Thus, both doses of Ta-1 modulated Lyt-1+/2+3+ ratios, with each dose affecting a different T-cell subset. The changes in the response to thyroglobulin are apparently exerted through the regulation of the functional T-cell subset balance.

Animals↗
PubMed1995

Dissimilarity between prostaglandin E1 and nitric oxide donors as potentiators of plasma exudation in the rabbit skin in vivo.

The ability of prostaglandin E1 (PGE1) and nitric oxide (NO) donor compounds such as sodium nitroprusside (SNP), glyceryl trinitrate (GTN), and 3-morpholino-sydnonimine (SIN-1) to modulate the histamine- and bradykinin-induced increase in microvascular permeability have been investigated in rabbit skin. The effect of the NO synthesis inhibitor N omega-nitro-L-arginine methyl ester (L-NAME) on the plasma exudation induced by histamine and bradykinin was also studied. Local edema formation was evaluated using [125I]human serum albumin. New Zealand white rabbits received an intravenous injection of [125I]human albumin followed immediately by the intradermal injection of edematogenic agents into the shaved dorsolateral skin. PGE1 (0.1 nmol/site) significantly potentiated both histamine- and bradykinin-induced edema. In contrast, SNP (0.4-400 nmol/site), SIN-1 (0.4-400 nmol/site), and GTN (0.4-40 nmol/site) did not affect the edematogenic response induced by either histamine or bradykinin. GTN (0.4-40 nmol/site) also had no effect on the increase in plasma exudation induced by histamine and bradykinin in the presence of PGE1. L-NAME (50-400 nmol/site, but not its enantiomer D-NAME, dose-dependently reduced the edema formation induced by a combination of either histamine or bradykinin with PGE1. This inhibition was significantly reversed by SNP (4-400 nmol/site) and by high doses (2.5 mumol/site) of L-arginine (but not by D-arginine). Our results thus demonstrate that PGE1, but not nitrovasodilators, can actually potentiate histamine- and bradykinin-induced edema in rabbit skin.(ABSTRACT TRUNCATED AT 250 WORDS)

Alprostadil↗
PubMed1995

Cytotoxic T lymphocyte and helper T cell responses following HIV polynucleotide vaccination.

Expression vectors encoding either HIV-1 gp160/rev, gp120, or rev alone were used for direct vaccination of mice and nonhuman primates. Each vaccine elicited long-lived (> 7 months) helper T cell responses in mice and monkeys as measured by in vitro proliferation of splenocytes following recombinant antigen treatment. Cytokine assays of the cell supernatants showed that approximately 100-fold more gamma-interferon than IL-4 was secreted during culture indicating that these vaccines elicited TH1-like responses. CD8+ CTL activities were also observed both in mice and rhesus. The gp120 and gp160/rev vaccines elicited antigen-specific antibodies, although these responses were more variable and lower magnitude for gp160/rev, and gp120 DNA-vaccinated African green monkeys had moderate levels of neutralizing antibodies. No antibodies were found against rev (an intracellular protein) with either rev vaccine. Similar antibody titers were obtained for gp120 by either intramuscular or intradermal injection although T cell responses were generally lower by intradermal route. These results indicate that DNA vaccines may provide a powerful means to elicit cellular and humoral immune responses against HIV.

AIDS Vaccines↗
PubMed2003

Factors affecting visualization rates of internal mammary sentinel nodes during lymphoscintigraphy.

UNLABELLED: There is great variation in the reported frequency of internal mammary (IM) sentinel node (SN) visualization. We observed a marked increase in our IM SN detection rate after 2 factors were changed simultaneously: depth of perilesional injection and dose. METHODS: A retrospective review of 82 consecutive patients (group 1) was compared with 61 consecutive patients (group 2) after changing the depth of perilesional injections and dose. Both groups had perilesional injections of (99m)Tc-sulfur colloid followed by intradermal injections at the areolar cutaneous junction. For group 2, activity was increased in all patients scheduled for next-day surgery. Group 2 had perilesional injections on top of, beside, and just below the estimated level of the tumor in an infiltrative manner, versus injections just on top of and beside the tumor as performed for group 1. RESULTS: The rates of IM SN visualization were 4.9% (4/82) for group 1 and 23.0% (14/61) for group 2 (P < 0.003). IM SNs were hotter in group 2 than in group 1. The total number of IM SNs detected per patient was also higher for group 2 than for group 1: 2.1 and 1.2, respectively. In group 2, patients with small breasts had an IM SN visualization rate of 46.2%; those with medium breasts, 21.1%; and those with large breasts, 0% (P < 0.017). In group 2, primary lesions located medially had a higher rate of IM SN visualization than did lesions located laterally: 38.9% (7/18) and 16.2% (6/37), respectively (P = 0.066). Dose was not a statistically significant factor within group 2 or group 1 when comparing IM SN visualization rate for doses above or below the mean or median. CONCLUSION: Modification of just these 2 factors resulted in a striking change in our IM SN detection rates. The injection depth was the most important factor. Breast size had a marked effect on the probability of detecting IM SNs. This suggests that the variation in detection rates reported in the literature could be at least partly dependent on variations in these factors, among others. Many surgeons do not routinely harvest IM SNs, but information about their presence can potentially alter treatment decisions.

Axilla↗
PubMed1986

Lymphocyte-induced angiogenesis: correlation with the metastatic incidence of two murine mammary adenocarcinomas.

Two tumor lines exhibiting different metastasizing capacity were studied with respect to the lymphocyte-induced angiogenesis (LIA). The MM3 line produce lung metastases with a high incidence. Conversely, the metastatic incidence of M3 line is much lower. Tumor cell suspensions were inoculated in BALB/c mice, 24 h later spleens were removed from the animals. Lymphocyte suspensions were made and then injected intradermally in syngeneic recipient mice. Five days after the injection of lymphocytes the vascular reaction was evaluated in the skin of the recipients by measuring the vessel density. The vascular response induced by lymphocytes from MM3-tumor bearing mice was increased when compared to that of M3-tumor-bearing mice. There appeared to be a positive correlation between metastasizing capacity and the lymphocyte-induced vascular response.

Adenocarcinoma↗
PubMedDate not supplied

Chronic inflammation induced by type I collagen in rats.

In this study we attempt to investigate if collagen type 1 from bovine Achilles tendon is capable of causing an inflammatory response. Thus, we immunized rats with a collagen emulsion injected in the right hind paw, via subplantar, and intradermally on the back. Seven days later, the rats were reimmunized by intradermal injection on the base of the tail with an identical emulsion. The inflammatory response was assessed by measuring the hind paw swelling of the rats every seven days for two months. We also analysed the levels of anticollagen antibodies in each rat during the experimental period. An indirect microELISA method was developed to determine collagen-specific-antibodies in rat serum. We observed a chronic inflammation in 41% of immunized rats. This pathology results in local swelling in the hind paw that remained for at least two months. This model was accompanied with the production of antibodies against collagen that appeared on day 14 after induction and remained during the study period (56 days). A positive correlation between swelling and antibody titre was observed.

Animals↗
PubMed1983

Guinea pig line 10 hepatocarcinoma model: characterization of monoclonal antibody and in vivo effect of unconjugated antibody and antibody conjugated to diphtheria toxin A chain.

Monoclonal antibodies were raised against the guinea pig line 10 (L10) hepatocarcinoma, and an IgG1-producing hybridoma (D3) was selected for further study. D3 is a true monoclonal antibody as demonstrated by two-dimensional gel electrophoresis. Radioimmunoassays on live cells revealed no cross-reactivity with normal tissues or with the line 1 hepatocarcinoma which was used as a control. Membrane immunofluorescence assays demonstrated similar specificity. Immunoperoxidase staining of cryostat sections of tumor and normal tissues of both adult animals and fetuses showed that the D3 monoclonal antibody reacted primarily with the L10 tumor, but some cross-reactivity with smooth muscle, placenta, fetal skeletal muscle, and fetal liver was also demonstrated. Radioimmunoprecipitation of detergent extracts of iodinated L10 cells showed that the antigen is present on the cell surface as a dimer of Mr 290,000 (unit size, Mr 148,000). Therapy studies with unconjugated D3 antibody demonstrated a minor dose-dependent effect on tumor growth. D3 antibody conjugated to the A chain of diphtheria toxin (10(-7) M) was cytotoxic to 100% of L10 cells in vitro. Animals treated with a single 1-mg i.v. injection of this immunoconjugate on Day 7 following the intradermal injection of 10(5) tumor cells demonstrated a highly significant inhibition of tumor growth compared to control animals and those treated with unconjugated antibody.

Animals↗