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PubMed1995

Cutaneous hypersensitivity induced in dogs and guinea-pigs by extracts of the tick Rhipicephalus sanguineus (Acari: Ixodidae).

The cutaneous hypersensitivity induced by Rhipicephalus sanguineus tick extract in dogs (natural host) and guinea-pigs (laboratory host) was evaluated. The left ear of infested and control (tick-bite naive) dogs and guinea-pigs was injected intradermally with an extract from unfed adult ticks and the right ear with phosphate buffered saline (PBS). Ear thickness variations were then measured after 10 min and 1, 2, 6, 18, 24, 48, 72 and 96 h post-injection. Results were expressed as percentual changes in the ear thickness in relation to pre-inoculation values. The final variation in ear thickness induced by the extract was given by subtracting, in each animal, the right ear percentual increase from that of the left ear. Guinea-pigs were tested at two different times following infestation and with two different doses of extract. Infested guinea-pigs from the three experiments developed an immediate (within the first 2 h post-inoculation) and a strong delayed reaction (24 h) to the extract. Dogs, unlike guinea-pigs, developed only a strong immediate reaction whereby an 80% increase in ear thickness was observed. Control animals, with the exception of one dog, did not develop any significant reaction to the extract. Only mild reactions were induced by PBS in the right ear of all animals. The correlation between the absence of a strong delayed type reaction to tick extract and the lack of resistance of the natural host to R. sanguineus tick is discussed.

Animals↗
PubMed1987

Epithelioid cell granuloma induction in the guinea pig by haptenated Mycobacterium leprae.

Fluorescein isothiocyanate (FITC)-conjugated Mycobacterium leprae (FITC-M. leprae) was injected intradermally into the ears of guinea pigs and granuloma formation in the draining postauricular lymph nodes was studied. At 2 weeks, there was an increase in weights and infiltration of the draining lymph nodes in half of the animals injected with FITC-M. leprae. At 5 weeks, there was a significant increase in the weights and infiltration of these draining lymph nodes in the guinea pigs injected with haptenated M. leprae compared with those injected with unconjugated M. leprae. At 5 weeks, there was also a significant increase in delayed type hypersensitivity responses to 25 micrograms purified protein derivative. Histologically, epithelioid cell granulomas were seen in these lymph nodes as early as 2 weeks when FITC-M. leprae was used as the source of antigen. Enhancement in the immunogenicity of M. leprae by conjugation with FITC has been postulated.

Animals↗
PubMed2004

Effects of gene delivery on collateral development in chronic hypoperfusion: diverse effects of angiopoietin-1 versus vascular endothelial growth factor.

OBJECTIVES: The aim of this research was to test the effects of vascular endothelial growth factor (VEGF)/angiopoietin-1 (Ang-1) on adult hypoperfused tissues. BACKGROUND: Angiopoietin-1 and VEGF act separately and synergistically in vascular development during embryogenesis. However, little is known regarding their relative roles in collateral development after chronic arterial obstruction and tissue ischemia in the adult. METHODS: Central and caudal ear arteries of 32 rabbits were ligated to induce ischemia. At two months, when flow was about 65% of pre-ligation values, we injected intradermally 10(9) plaque-forming unit adenovirus with the following transgenes: Ang-1, VEGF, or a combination of both. Ear perfusion was followed up for four weeks, and vessel leakage was assessed by Evens Blue test. RESULTS: Before injection, flow was 65% of baseline, and endogenous VEGF levels in ischemic tissue were increased. Adenovirus-encoding VEGF gene (Ad.VEGF) at one week caused a visible inflammatory response associated with a 24% flow increase (p = 0.018). Adenovirus-encoding Ang-1 gene (Ad.Ang-1) increased flow 22% (p = 0.004) with no visible inflammation; Ad.VEGF caused three times as much vessel leakage as Ad.Ang-1 (142.5 +/- 38 vs. 49.5 +/- 9.8 ng Evens Blue/mg tissue; p < 0.001). However, at four weeks, compared with baseline, VEGF decreased flow 18% (p = 0.004), whereas Ang-1 increased tissue perfusion 26% (p < 0.001). This effect was abolished when Ad.Ang-1 was injected with soluble VEGF receptor [Ad.Flt(1-3)-Fc], which blocks VEGF-dependent signaling. Exogenous Ang-1 did not increase perfusion in a normally perfused ear, in which endogenous VEGF is not expressed. CONCLUSIONS: Exogenous Ang-1 enhances perfusion in hypoperfused tissues only in the presence of increased levels of endogenous VEGF. Overexpression of VEGF, however, after causing an inflammatory response, does not improve collateral blood flow.

Adenoviridae↗
PubMed2000

Antinociceptive activity of ricinoleic acid, a capsaicin-like compound devoid of pungent properties.

The antinociceptive effect of ricinoleic acid ([R-(Z)]-12-hydroxy-9-octadecenoic acid) in comparison with capsaicin (trans-8-methyl-N-vanillyl-6-nonenamide) has been investigated in several "in vivo" tests. Acute topical application of capsaicin, but not ricinoleic acid, produced by itself an hyperalgesic effect detected as a decrease in paw withdrawal latency in response to a painful (heat) stimulus in mice. Capsaicin, but not ricinoleic acid at any dose tested, showed an irritant effect in the wiping test in guinea pig conjunctiva after local application and in the paw licking test in mice after intradermal injection. Whereas acute application of ricinoleic acid or capsaicin decreased paw withdrawal latency to heat in the presence of a pre-existing inflammation (injection of carrageenan in the mouse paw), the repeated local treatment for 8 days with either compounds markedly increased paw withdrawal latency. In a chronic model of inflammation (complete Freund's adjuvant arthritis in mice), the repeated topical and intradermal treatments with both ricinoleic acid and capsaicin increased paw withdrawal latency to heat, the antinociceptive effect of ricinoleic acid being more persistent than that of capsaicin. Antinociceptive effect of 8 days of treatment with ricinoleic acid and capsaicin was observed in acetic acid-induced writhing in mice, capsaicin-induced foot licking in mice and capsaicin-induced wiping movements in guinea pig conjunctiva. A decrease of substance P tissue levels in the mouse paw was found after repeated treatment with ricinoleic acid. In conclusion, ricinoleic acid seems to be a new antinociceptive agent lacking the pungent and acute hyperalgesic properties of capsaicin.

Administration, Topical↗
PubMed1994

Mild hypothermia during isoflurane anesthesia decreases resistance to E. coli dermal infection in guinea pigs.

Small changes in core temperature profoundly alter cutaneous blood flow, a major factor influencing resistance to wound infection. Furthermore, when measured in vitro, various immune functions are temperature dependent in the physiological range. Accordingly, we tested the hypothesis that mild hypothermia impairs and mild hyperthermia improves resistance to dermal infections. Thirty-two guinea pigs were anesthetized for 6 h using 1.5% (1.25 MAC) inspired isoflurane. Their core temperatures were maintained at either 39 degrees C (normal for guinea pigs, n = 11), 36 degrees C (n = 12), or 41 degrees C (n = 9). One h after induction of anesthesia, 2 x 10(8) E. coli were injected intradermally with a 26-g needle at eight sites on each animal's back. Core temperatures were not controlled after recovery from anesthesia, and animals in each group were maintained in the same environment. Twenty-four h after injection, the area of induration surrounding each injection site was measured. This is a standard test of resistance to wound infection. Values were compared using one-way ANOVA and Scheffé's S tests. Results are presented as means +/- standard deviations; differences were considered significant when P < 0.05. Areas of inflammation on the hypothermic animals were significantly larger (48 +/- 10 mm2) than those on normothermic (36 +/- 10 mm2) or hyperthermic (37 +/- 6 mm2) animals. These data suggest that mild hypothermia during anesthesia significantly impairs resistance to dermal infection. In contrast, mild hyperthermia does not appear to be protective.

Anesthesia, Inhalation↗
PubMed1988

Extracellular amino acid concentrations in the dorsal spinal cord of freely moving rats following veratridine and nociceptive stimulation.

In vivo microdialysis was used to sample extracellular concentrations of amino acids in the dorsal lumbar spinal cord of freely moving rats. Changes in the extracellular concentrations of amino acids were measured in response to infusion of veratridine (180 microM), a sodium channel activator, as well as during acute noxious stimulation by an injection of 5% formalin into the metatarsal region of the hindleg. Veratridine produced a tetrodotoxin (TTX)-sensitive increase in the extracellular concentration of Glu. Concentrations of Asp, taurine, Ala, Asn, and Gly were not significantly elevated following veratridine stimulation. Intradermal injection of formalin produced a TTX-sensitive increase in Asp concentration and a non-TTX-sensitive increase in Glu concentration. These data support the hypothesis that Glu and Asp are dorsal horn neurotransmitters involved in nociception.

Amino Acids↗
PubMed1965

[Studies on the intradermal test of Clonorchis peptide(CPT) Antigen 1. Preliminary report]

Polypeptide antigen as isolated from adult worms of Clonorchis sinensis by means of diluted HCl-extraction method. Preliminary intradermal examination with those polypeptide antigen(CPT) were performed on patients of clonorchiasis, paragonimiasis and metagonimiasis and the result of the intradermal test with CPT antigen were compared with those of Clonorchis protein fraction(CPF) and Veronal buffer saline extraction (VBS) antigen. The result of the examination were summerized as follows: 1. When 0.05 ml (1 microgram) of CPT was injected intradermally, the positive intradermal reaction appeared in 42.7 per cent (37 of 82) on the patients of clonorchiasis. 2. The sizes of the wheal and erythema with CPT antigen was smaller than that of CPF and VBS antigen. 3. Intradermal reaction with CPT antigen was carried out for other parasitic diseases, such as paragonimiasis and metagonimiasis. The result indicated that CPT antigen may be showed no cross reactions. 4. Clonorchiasis pateints who with negative reaction to CPT antigen, reacted negatively to CPF and VBS antigens.

Journal Article↗
PubMed1990

Evaluation of cell-mediated, cutaneous basophil hypersensitivity in young chickens by an interdigital skin test.

An evaluation was made of an interdigital skin test to assess cutaneous basophil hypersensitivity (CBH), mediated by thymic (T) cells, in chickens less than 2 wk of age. Hubbard-by-Hubbard roosters 3, 5, 7, 10 and 14 days of age were injected intradermally in the interdigital skin between Digits 3 and 4 of the right foot with 100 micrograms or 200 micrograms of phytohemagglutinin-P (PHA-P) in .10 mL of physiological saline solution (PSS). The left foot was injected with PSS and served as a control. The CBH response elicited by the PHA-P was evaluated by determining interdigital skin thickness before injection and at 12 and 24 h after injection. The effect of treatment with the T-cell suppressing drugs dexamethasone and cyclosporin on the CBH response was determined in chickens treated daily for 4 or 5 days prior to skin testing at 5 or 10 days of age. Significant increases (P less than .005) of .64 to .88 mm occurred in the mean, interdigital skin thickness for all age groups of chickens at 12 and 24 h after injection of 100 or 200 micrograms of PHA-P. The PHA-P-elicited. CBH response was significantly suppressed by treatment with dexamethasone (P less than .005) or cyclosporin (P less than .05). The results indicated that the interdigital skin test is a rapid and simple means of assessing normal and suppressed immune responses in chickens at 3 to 14 days of age.

Animals↗
PubMed1978

Investigation of the reputed inhibition of the Prausnitz-Küstner reaction by the pentapeptide Asp-Ser-Asp-Pro-Arg.

An investigation has been made of the inhibitory activity of the so-called Hamburger pentapeptide, Asp-Ser-Asp-Pro-Arg, in both the human Prausnitz-Küstner and baboon passive cutaneous anaphylaxis systems. With a dose of 6 microgram peptide, administered intradermally 24 h prior to diluted allergic serum from a horse-dandruff-sensitive individual, partial inhibition of wealing was observed in three different normal recipients following the pricking in of specific allergen. In one of the recipients, inhibition of flaring was observed at test sites subsequently challenged with timothy grass pollen allergen. A similar dose of pentapeptide, again injected intradermally 24 h prior to passive sensitisation, was shown to inhibit extravasation in a normal baboon on subsequent challenge with horse dandruff (but not grass pollen) allergen by subcutaneous injection into the upper arm. In vivo tests performed in the forearms of grass-pollen-sensitive individuals failed to provide convincing evidence of displacement of mast-cell-bound IgE antibody by the pentapeptide.

Animals↗
PubMed2001

[Sentinel node involvement in T0-T1 breast cancers].

STUDY AIM: Determination of axillary lymph node status is crucial in diagnosis of early breast cancer. However thanks to an early diagnosis, an increasing number of axillary lymph node dissections are free of disease. This raises questions about the need for this procedure. The study aim was to report an experience with lymphadenectomy and sentinel node mapping in patients with T0-T1 carcinoma of the breast. METHODS: Between November 1997 and December 1999, 84 consecutive women (T0-T1 N0 according to the 1987 UICC classification) with recently diagnosed breast cancer, were included in this study for identification of the sentinel lymph node (SLN). The SLN was removed and submitted for histological examination. All patients underwent axillary dissection; nodes from levels I and II (Berg's classification) were excised and submitted to histological examination. RESULTS: The average tumor diameter was 12.7 mm (range, 3 to 25 mm). The lymphatic mapping technique was obtained after injection of the isotope into the breast around the tumor in 53/84 patients: the sentinel lymph node was the only positive node in 10 patients and it was positive in 5 patients with other axillary nodes. In 15/84 patients, an intradermal injection of blue dye was used; two sentinel nodes were positive and one falsely negative. In 16/84 patients, an interdermal injection of blue dye was used to make up for. In this study, the sentinel node was positive in three patients and falsely negative in one patient. The discrepancy was due to an important involvement of an axillary area excluded from the lymphatic channels. 22/84 patients (26%) had a metastatic spread to the axillary nodes. 30/84 patients had also an isotopic captation in another lymph node group (internal mammary). CONCLUSION: This study confirms that lymphatic mapping is technically possible in the patients with T0-T1 breast cancer and that the histological characteristics of the sentinel node probably reflect the histological characteristics of the rest of the axillary lymph nodes, but do not provide any information about the other lymph node sites.

Adult↗
PubMed1995

Repeated exposure to silicone gel can induce delayed hypersensitivity.

The possible immunologic reactivity of silicone gel remains speculative and controversial. In this laboratory, a quantitative lymphocyte localization assay has been developed and well studied using pure lymphocytes collected by the technique of lymph vessel cannulation in sheep. The kinetics of antigen-specific immune responses (e.g., tuberculin reaction) in this model are well described and accepted. Using the known parameters regarding the response to purified protein derivative and the classic adjuvant Freund's Complete Adjuvant, this study was designed to identify the possible antigen-specific immunologic response, in the form of delayed-type hypersensitivity, after repeated exposure to silicone gel. Pure lymphocytes were collected by cannulating the efferent vessel of a subcutaneous lymph node in four groups of primed sheep which, 30 days previously, had received intradermal injections of 0.9% saline (negative controls; n = 6), Freund's Complete Adjuvant only (positive controls; n = 6), silicone gel (n = 7), or Freund's Complete Adjuvant homogenized with silicone gel (n = 7) in an attempt to induce sensitization. Multiple (1040) intradermal skin tests were performed using silicone gel, purified protein derivative, and 0.9% saline. After the skin lesions had developed for 48 hours, 5 x 10(8) lymphocytes were labeled in vitro with indium-111, returned intravenously, and allowed to circulate for 3 hours. Sheep were euthanized, the skin lesions were removed, and the radioactivity was counted in a gamma spectrometer. The radioactivity in each skin lesion is considered a measure of lymphocyte accumulation. The occurrence of augmented accumulation after reexposure to an antigen is a hallmark of delayed-type hypersensitivity. The purified protein derivative and saline lesions functioned as positive and negative controls, with counts per minute (cpm +/- standard error) of 2404 +/- 478 (Freund's Complete Adjuvant group) and 149 +/- 21 (saline group), respectively. Significantly greater (p = 0.0021) radioactivity was found in the silicone gel sites (310 +/- 35) in the silicone gel-primed group and the Freund's Complete Adjuvant plus silicone gel group (453 +/- 44; p = 0.0004) than in normal skin in each group. These data suggest that it may be possible to induce an antigen-specific lymphocyte-mediated response to silicone gel.

Animals↗
PubMed1976

Phytohaemagglutinin (PHA) skin test in predicting prognosis in cancer patients.

The response to intradermal injection of 1 mug of purified PHA was found to correlate with survival of patients with advanced solid tumours treated by chemotherapy, with responders surviving longer than non-responders. In patients with prolonged survival, the reaction was enhanced. Negative intradermal response to PHA was found to be less common in cancer patients than negative responses to tuberculin or streptokinase, and was more easily reversed by successful chemotherapy.

Adult↗
PubMed1999

Strain-dependent induction of allergic sensitization caused by peanut allergen DNA immunization in mice.

To investigate the potential application of allergen gene immunization in the modulation of food allergy, C3H/HeSn (C3H) mice received i.m. injections of pAra h2 plasmid DNA encoding one of the major peanut allergens, Ara h2. Three weeks following pDNA immunization, serum Ara h2-specific IgG2a, IgG1, but not IgE, were increased significantly in a dose-dependent manner. IgG1 was 30-fold higher in multiply compared with singly immunized mice. Ara h2 or peanut protein injection of immunized mice induced anaphylactic reactions, which were more severe in multiply immunized mice. Heat-inactivated immune serum induced passive cutaneous anaphylaxis, suggesting that anaphylaxis in C3H mice was mediated by IgG1. IgG1 responses were also induced by intradermal injection of pAra h2, and by i.m. injection of pOMC, the plasmid DNA encoding the major egg allergen protein, ovomucoid. To elucidate whether the pDNA immunization-induced anaphylaxis was a strain-dependent phenomenon, AKR/J and BALB/c mice also received multiple i.m. pAra h2 immunizations. Injection of peanut protein into these strains at weeks 3 or 5 following immunization did not induce reactions. Although IgG2a was increased significantly from week 2 in AKR/J mice and from week 4 in BALB/c mice and remained elevated for at least 6 wk, no IgG1 or IgE was detected. These results indicate that the type of immune responses to pDNA immunization in mice is strain dependent. Consequently, models for studying human allergen gene immunization require careful selection of suitable strains. In addition, this suggests that similar interindividual variation is likely in humans.

Allergens↗
PubMed1988

Efficacy of a cell-mediated reaction to the purified protein derivative of tuberculin in the disposal of Mycobacterium leprae from human skin.

The purpose of this study was to evaluate the effects of a delayed-type cell-mediated immune response to Mycobacterium tuberculosis antigen on the Mycobacterium leprae load in the skin of leprosy patients. Twelve patients with the lepromatous form of leprosy have been injected intradermally with 5 units of the purified protein derivative of tuberculin (PPD). Ten individuals responded with areas of induration ranging from 12 to 21 mm in diameter, and two were unresponsive (less than 10 mm). Twenty-one days thereafter, the injected and control sites were biopsied, and the histology, number of acid-fast bacilli, nature and phenotype of the emigrant cells, and ultrastructural characteristics of the lesions were evaluated. Eight of the 10 responding patients showed reductions in the number of acid-fast bacilli by factors ranging from 5 to 10,000. Two responders and both nonresponders exhibited no discernible decline in the number of organisms. The reduction in bacillary load was correlated with an intense mononuclear cell infiltrate, the maintenance of a high CD4+ T-cell/CD8+ T-cell ratio, the formation of granulomata, and the extensive destruction of previously parasitized macrophages.

Antigens, Bacterial↗
PubMed1982

[General pharmacology of T-1982, a new cephamycin antibiotic].

General pharmacological studies on T-1982 produced the following results. On central nervous system, subcutaneous injection of T-1982 at dose of 2,000 mg/kg hastened the onset of pentetrazole-induced tonic extensor in mice. T-1982 had no effect on spontaneous motor activity, pentobarbital hypnosis, body temperature or EEG in mice or rabbits, and also did not show motor incoordinate, anticonvulsive or analgesic activity in mice at intravenous doses of 250--1,000 mg/kg or subcutaneous doses of 500--2,000 mg/kg. On motor and sensory nervous systems, no effect of T-1982 was noted on spinal reflex, neuromuscular junction, conduction anesthesia or surface anesthesia in rats or rabbits. On respiratory, cardiovascular and autonomic nervous systems, T-1982 caused transient increase of respiratory rate, slight hypotension and transient increase of femoral blood flow in dogs at intravenous doses of 250--1,000 mg/kg. However, it caused a slight hypertensive tendency in rabbits. Heart rate and ECG in dogs or rabbits, blood pressure response to epinephrine, isoproterenol, acetylcholine or histamine in dogs, nictitating membrane in cats and pupil size in mice were not affected after intravenous injection of T-1982. No effect was found on isolated guinea pig atrium or rabbit descending aorta following T-1982 application. On renal function in rats, T-1982 caused an increase of PSP excretion but had no effect on urine volume or electrolytes excretion at intravenous doses of 250--1,000 mg/kg. T-1982 prolonged bleeding time in mice at intravenous doses of 500--1,000 mg/kg, but did not show hemolytic property and inhibitory activity on blood coagulation or platelet aggregation in vitro experiments. Spontaneous movement and tone of isolated stomach, ileum, colon, uterus, vas deferens or trachea and acetylcholine-, histamine-, nicotine- or barium chloride-induced contraction of ileum were not affected following T-1982 application. Intestinal propulsion of barium meal in mice, gastric secretion and carrageenin-induced edema in rats were not affected after intravenous injection of T-1982. T-1982 increased bile secretion in rats dose-dependently at intravenous doses of 31.3--125 mg/kg. The local irritative activity of T-1982 in rats was slightly milder than cefoxitin and moderately milder than cefmetazole after intradermal injection. In conclusion, these results suggest that T-1982 would not cause any adverse effects at its estimated clinical doses of 10--20 mg/kg (500--1,000 mg/man).

Animals↗
PubMed2003

Injection of genetically engineered fibroblasts corrects regenerated human epidermolysis bullosa skin tissue.

Current therapeutic strategies for genetic skin disorders rely on the complex process of grafting genetically engineered tissue to recipient wound beds. Because fibroblasts synthesize and secrete extracellular matrix, we explored their utility in recessive dystrophic epidermolysis bullosa (RDEB), a blistering disease due to defective extracellular type VII collagen. Intradermal injection of RDEB fibroblasts overexpressing type VII collagen into intact RDEB skin stably restored correctly localized type VII collagen expression in vivo and normalized hallmark RDEB disease features, including subepidermal blistering and anchoring fibril defects. This article was published online in advance of the print edition. The date of publication is available from the JCI website, http://www.jci.org.

Animals↗
PubMed1991

Evaluation of relationship between plasma fibrinogen concentration and tuberculin testing in red deer.

After intradermal injection of bovine purified derivative (PPD), increases in plasma fibrinogen concentration and plasma viscosity developed in red deer (Cervus elaphus) with a history of tuberculosis caused by Mycobacterium bovis. Serum haptoglobin concentrations were also found to increase under similar circumstances. The increases were reproducible and did not appear to be related to mustering, stress, or the handling associated with injection of PPD. A significant (P less than 0.05) direct relationship was found between the increase in plasma fibrinogen concentration and various markers of bovine tuberculosis infection, such as stimulation of lymphocyte transformation in response to bovine PPD and the diameter of intradermal tuberculin skin test reactions. A stronger correlation (P less than 0.01) was found with the volume of intradermal tuberculin skin test reactivity, and the strongest correlation (P less than 0.001) was with the presence of circulating antibovine PPD antibody.

Animals↗
PubMed1978

Suppression of growth of guinea pig line-10 hepatocarcinoma. I. Effect of simultaneous administration of tumor cells and antibodies from normal rabbits.

Suppression of growth of the line-10 hepatocarcinoma in strain-2 guinea pigs occurred when line-10 cells were injected intradermally together with sera or immunoglobulins derived from normal rabbits. A significant number of animals were resistant to subsequent rechallenge with tumor cells. This immunity was specific, depended on contact of immunoglobulins with tumor cells and on the concentration of immunoglobulins. Repeated injections acted as potent vaccines and resulted in the development of immunity in 84.6% of recipients. Fc receptors were not detected on line-10 cells. Antibodies reacting with line-10 cell unique antigens as well as with antigens common to line-10, line-1 and normal guinea pig spleen cells were found in NRS. Injection of line-10 cells together with rabbit immunoglobulins from which antibodies reacting with antigens derived from line-10 cells had been removed did not result in tumor suppression. The specific antigen(s) recognized by antibodies that suppressed growth of the line-10 tumor in vivo was not determined.

Animals↗