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PubMed2005

Technetium labeling of dextran incorporating cysteamine as a ligand.

INTRODUCTION: Technetium-99m-labeled dextran is a useful imaging agent for procedures such as angiocardiography and lymphoscintigraphy. To improve the availability of 99mTc-labeled dextran, we designed a cysteamine ligand system for dextran labeling. METHODS: Cysteamine derivatized dextran was synthesized as follows. Dextran was oxidized with sodium periodate, coupled with cysteamine and reduced with sodium borohydride to provide the desired amine ligand. The cysteamine-dextran conjugate was then labeled with reduced 99mTc. Whole-body scintigraphy and biodistribution were examined following injection of the 99mTc-labeled cysteamine-conjugated dextran (99mTc-cysteamine-dextran) in ICR mice. Lymphoscintigraphy was performed after intradermal injection of 99mTc-cysteamine-dextran in SD rats. RESULTS: The cysteamine-derived dextran was easily labeled with reduced 99mTc in greater than 96% yield. 99mTc-cysteamine-dextran has a higher chelation stability against diethylenetriamine pentaacetic acid (DTPA) than the 99mTc-dextran. Axillary lymph nodes were clearly visible after intradermal injection of 99mTc-cysteamine-dextran in rats. CONCLUSION: These results suggest that 99mTc-cysteamine-dextran is available for lymphoscintigraphy. This methodology could expand the usage of 99mTc-labeled dextran, particularly for diagnostic purposes.

Animals↗
PubMed1999

Migration of human dendritic cells after injection in patients with metastatic malignancies.

Present clinical studies of active immunotherapy for malignancies using dendritic cells (DCs) require elucidation of the sites where DCs localize after injection. We evaluated the pattern of distribution of in vitro-generated, antigen-loaded, human DCs labeled with indium-111 oxyquinoline after i.v., s.c., and intradermal injection. Whereas the DCs injected i.v. localized in the lungs and then redistributed to the liver, spleen, and bone marrow, they were not detected in lymph nodes or tumors. A small percentage of DCs injected intradermally migrated rapidly to the regional lymphatics in some individuals. No lymph node activity was detected after s.c. injection. Our results demonstrate that DC distribution to sites of lymphoid tissue is dramatically affected by the mode of administration.

Cell Movement↗
PubMed1990

Fluorescence microscopic demonstration and demarcation of doxorubicin extravasation. Experimental and clinical studies.

Extravasation of doxorubicin and its derivative epirubicin, cause severe progressive tissue necrosis that requires early excision of all affected tissues. Doxorubicin and epirubicin are autofluorescent and this characteristic can be used to demonstrate and delineate extravasation by the aid of fluorescence microscopic study. In a rat model doxorubicin was injected intradermally in declining concentrations and the minimal detectable concentration was found to be 0.02 mg/ml. Skin necrosis developed in rats injected with doxorubicin concentrations ranging from 0.02 mg/ml to 2 mg/ml. Clinically, fluorescence microscopic analysis of frozen sections was used in eight patients to assess whether doxorubicin or epirubicin extravasation had taken place. When all fluorescing tissue was removed, no necrosis ensued, but in one patient, where a slightly fluorescing area was ignored, necrosis developed later and excision of the tissue had to be performed.

Adult↗
PubMed1998

Intercellular adhesion molecule-1 expression in adjuvant arthritis in rats: inhibition by kappa-opioid agonist but not by NSAID.

OBJECTIVE: To quantify intercellular adhesion molecule-1 (ICAM-1) expression in normal and adjuvant arthritic rats and to determine the extent to which ICAM-1 expression in arthritic animals is altered by treatment with a prototype nonsteroidal antiinflammatory drug (NSAID) and a kappa-opioid agonist. METHODS: Unilateral hind paw inflammation was induced by intradermal injection of Freund's complete adjuvant (FCA) into the right hind paw of female Lewis rats. Polyarthritis was induced by intradermal injection of FCA into the base of the tail of female dark Agouti rats. The NSAID naproxen [5 mg/kg intraperitoneally (ip)] or the kappa-opioid PD117302 (15 mg/kg ip) was administered twice daily throughout the experiment (21 days). ICAM-1 expression was quantified using monoclonal antibodies against rat ICAM-1 that bind to the endothelium in proportion to the degree of adhesion molecule expression. RESULTS: ICAM-1 expression was significantly upregulated in the joints of affected limbs of animals with both unilateral hind paw inflammation and polyarthritis. In animals treated with PD 117302 and naproxen there was a significant attenuation of arthritis; however, only treatment with PD117302 was able to significantly inhibit the upregulation of ICAM-1 expression in arthritic joints. CONCLUSION: ICAM-1 expression is upregulated in experimental arthritis. It appears that the kappa-opioid PD117302, but not the NSAID naproxen, inhibits the upregulation of ICAM-1 in arthritic joints, suggesting these agents act via different mechanisms. The ability of the kappa-agonist, PD117302, to inhibit both the inflammation and upregulation of ICAM-1 in arthritic joints emphasizes the potential of kappa-agonists as antiarthritic agents.

Animals↗
PubMed1992

Peripheral and central mediators of itch.

Since the majority of itching skin diseases are inflammatory or allergic, it has been assumed that release or activation of inflammatory mediators, stimulating the itch receptors, play an essential role in the pathophysiology of itch. In this review some of the possible mediators are discussed. Histamine induces itch upon intradermal injection, but urticaria is the only itching dermatosis which is significantly relieved by antihistamines. Serotonin is much weaker than histamine in provoking itch upon intradermal injection. Serotonin acting in synergism with prostaglandins may cause itch in polycythaemia vera. Neuropeptides release histamine from skin mast cells, but it remains to be determined whether neurogenic peptides are responsible for clinical pruritus. Prostaglandins enhance pruritus induced by intradermal histamine (and serotonin) but are weak pruritogens per se. Lymphocytes are present in many itching skin diseases and it could be assumed that lymphokines are involved in the pathogenesis of itch. Supporting this theory is the finding that ciclosporin A, an inhibitor of lymphokine production, reduces itch in atopic dermatitis. Central mechanisms are essentially unknown, but there are indications that opioid peptides might be involved in the central transmission of itch.

Eicosanoids↗
PubMed2006

Experimental induction of nocardiosis in yellowtail, Seriola quinqueradiata Temminck and Schlegel by artificial challenge.

Challenge methods for inducing nocardiosis, caused by Nocardia seriolae in yellowtail, Seriola quinqueradiata, were evaluated. The first method involved intraperitoneal injection of 1.1 x 10(2), x 10(3) and x 10(4) cfu of N. seriolae; the second was by bath immersion with three different concentrations of bacterial suspension for 10 min; the third was by intradermal injection using a multipuncture device; the fourth was by oral administration using a tube; the fifth was based on the co-habitation of uninfected fish with others that had been artificially infected, i.e. intraperitoneally or by immersion for 10 min. The LD(50) values for the infection trials were 1.9 x 10(2) (intraperitoneal injection), 1.5 x 10(4) mL(-1) (immersion), 4.3 x 10(6) (intradermal injection) and 1.7 x 10(7) (oral administration). In the co-habitation challenge, mortalities were 70% and 50% in groups of non-infected fish mixed with fish infected by the i.p. injection and immersion methods, respectively. Fish challenged by intraperitoneal injection developed adhesions. Fish challenged by other methods did not show any gross clinical signs and moribund fish were similar to natural infection.

Animals↗
PubMed1990

Encephalitogenicity for rats of myelin basic protein without the aid of water-in-oil emulsions.

The induction of experimental allergic encephalomyelitis (EAE) with purified myelin basic protein (MBP) has, heretofore, required its incorporation in a water-in-oil emulsion or adsorption on particulate adjuvants. In the present work, the absorption of a saline solution of MBP from the peritoneal cavity into the mediastinal lymph nodes was increased by giving repeated inoculations or by pretreating rats with a peritoneal irritant. Under these conditions, the only adjuvant needed for production of EAE was aqueous pertussis vaccine which was injected separately a few hours or one day after the MBP. Pertussis vaccine was also necessary for production of EAE with intradermal injection of aqueous MBP. By injecting the aqueous MBP directly into pre-enlarged popliteal lymph nodes, it was possible to produce EAE without the pertussis vaccine. Thus, EAE can be induced in rats using MBP without the addition of Freund's adjuvant or pertussis vaccine.

Animals↗
PubMed1969

Allergenicity of mycobacterial ribosomal and ribonucleic acid preparations in mice and guinea pigs.

Guinea pigs were injected subcutaneously with mycobacterial ribosomal fraction incorporated in Freund's incomplete adjuvant and tested 6 and 12 weeks later by the intradermal injection of 0.5 mug (25 TU) of Purified Protein Derivative. No evidence of delayed-type hypersensitivity could be detected in these animals, although large necrotic reactions were obtained in guinea pigs sensitized with living, attenuated mycobacterial cells. Mice also were vaccinated by the intraperitoneal injection of mycobacterial ribosomal fraction or ribonucleic acid (RNA) and tested for sensitivity to tuberculin at various subsequent times. No evidence of true tuberculin hypersensitivity could be detected at any time, although what appeared to be small Arthus type reactions were seen in mice given the largest vaccinating doses. Attempts to recall tuberculin sensitivity in vaccinated mice by the intravenous injection, 4 weeks after vaccination of living cells, of either the virulent or attenuated mycobacterial strains were unsuccessful. Instead, when the virulent cells were injected, a suppression of footpad reactivity was noted in animals made sensitive to tuberculin by the previous intraperitoneal injection of viable attenuated mycobacterial cells. Both guinea pigs and mice, vaccinated as described above, were also skin tested or footpad tested, respectively, with 2 mug of the ribosomal fraction or RNA used for vaccination. No evidence of true tuberculin hypersensitivity could be obtained; instead, in guinea pig skin very small dermonecrotic areas were noted, and in mice swelling and redness of the footpad occurred to an equal extent in both vaccinated and nonvaccinated mice. The possible role of tuberculin hypersensitivity in acquired immunity to tuberculosis is discussed, and the conclusion is reached that its part, if any, is minor.

Allergens↗
PubMed1995

Airway responses following intradermal sensitization to different types of allergens: ovalbumin, trimellitic anhydride and Dermatophagoides farinae.

Sensitization of guinea pigs by intradermal injections of the occupational allergen trimellitic anhydride (TMA) in oily vehicle has been shown to be very reproducible. We studied the effect of intradermal sensitization with ovalbumin (OA) in oily vehicle on immune and airway responses in guinea pigs. We also compared airway responses to trimellitic anhydride or Dermatophagoides farinae (DF; mite) with those to OA in guinea pigs intradermally sensitized to respective allergens. Three to four weeks after sensitization, the animals were challenged with intratracheal instillation of these allergens. Intradermal injections with OA developed dose-dependently specific IgG1 antibodies to OA demonstrated by ELISA. In animals sensitized with different doses of OA in corn oil vehicle, a challenge with OA induced a reversely dose-dependent airflow obstruction and airway plasma exudation. In contrast, animals sensitized with OA in saline vehicle had dose-dependent airway responses to OA. Challenge with OA caused an immediate peak and subsequently persistent airflow obstruction, whereas this response to either TMA guinea pig serum albumin or Df was slowly progressive in animals sensitized to respective allergens. The animals sensitized to TMA or Df may show a different profile of airway responses following the challenge compared to OA. Intradermal sensitization may be a valuable method of sensitization for the development of an animal model of airway allergy to different types of allergens, including chemicals or mites.

Allergens↗
PubMed1984

Inhibition of bacterial clearance in the guinea pig by fluid-phase C3b.

The large-molecular-weight activation product C3b of C3, the third component of complement, loses its opsonic properties when it is free in solution (fluid phase) rather than attached to a particle, eg, a bacterial cell. Human fluid-phase C3b, which has been shown in vitro to be an inhibitor of the bactericidal function of human neutrophils, was injected intradermally or intraperitoneally into guinea pigs along with various species of bacteria, and the clearance rate of the bacteria was measured. The injected C3b caused a large increase in the time required for the clearance of the bacteria. The inhibition by C3b of clearance of the injected bacteria was dependent on the bacterial species; bacteria that depended strongly on complement for opsonophagocytosis was cleared at a much lower rate in the presence of fluid-phase C3b than were bacteria that did not have the corresponding dependence on complement.

Animals↗
PubMed1987

Application of thermography to the evaluation of the histamine skin test in man.

The degree of skin flare 10 min after intradermal injection of histamine (0.06-2 micrograms in 0.1 ml saline) was evaluated both visually and by thermography in 6 healthy male volunteers. Intradermally injected histamine increased the skin flare area in a dose-dependent manner. There was a good exponential correlation (r = 0.963) between the flare area evaluated visually and that evaluated by thermography, indicating that the degree of underestimation of the flare area by visual inspection increased with increasing flare size. This could be due to the fact that the rather irregular shape of a flare was rounded by the eyes. Suppressive effects of an antihistamine, terfenadine (20, 40, 60, and 120 mg, p.o.), on the flare caused by histamine (2 micrograms/0.1 ml saline) were evaluated in a similar way in a single-blind, crossover controlled trial using 15 healthy male volunteers. The histamine skin test was evaluated both visually and by thermography before and 4 hr after administration of placebo or each of 4 doses of terfenadine. Terfenadine (20, 40, and 60 mg) suppressed the skin flare in a dose-dependent manner, but 60 and 120 mg had almost the same effect. In all cases the flare area evaluated visually was smaller than that obtained by the thermography, although the degrees of reduction of flare area by the drug were similar by both methods when expressed as a percentage of placebo. In conclusion, thermography could serve as a tool for the objective evaluation of the histamine skin test.

Adult↗
PubMed1990

Solar urticaria: studies on mechanisms of tolerance.

The mechanisms by which tolerance is induced in solar urticaria were investigated in two patients whose eruptions were induced by wavelengths in the range 320-455 nm in one patient and 400-495 nm in another. Tolerance to radiation was induced by repeated exposures of the skin to the eliciting wavelengths of light. Weal and flare responses to intradermal injections of histamine and the histamine-releasing agent (codeine) were unaltered in the tolerant skin when compared with adjacent normal skin. Intradermal injection of in vitro irradiated serum or plasma from the patients induced an urticarial reaction in the unexposed skin but not in tolerant skin and repeated injections induced tolerance to the eliciting radiation. The results suggest that tolerance is not due to exhaustion of the photoallergen in the skin, or to an increase of the mast-cell degranulation threshold caused by exposure to ultraviolet radiation, or mast-cell mediator depletion, or histamine tachyphylaxis. It is likely that binding sites of IgE on mast cells remain occupied by the photoallergen during the state of tolerance, and that histamine release from mast cells is blocked.

Aged↗
PubMed1983

Half-life of murine IgE antibodies in the mouse.

The half-life of intravenously injected murine anti-DNP IgE antibody was determined in recipient mice by sequential bleeding and was found to be 12 h. Intradermally injected IgE antibody persisted much longer, the half-life was 6 days.

Animals↗
PubMed2002

Distinct temporal patterns of macrophage-inflammatory protein-2 and KC chemokine gene expression in surgical injury.

In the present study the regulation of CXC chemokine expression was evaluated in full-thickness abdominal wounds in mice. During the first 24 h after injury, IL-1alphabeta, KC, macrophage-inflammatory protein (MIP)-2, and monocyte chemoattractant protein-1 were the predominant cytokines and chemokines produced; TNF-alpha was not detected. Chemokine mRNA expression and protein secretion occurred in two temporal stages. The first, which reached a maximum at 6 h, was associated with high levels of IL-1alpha and KC and low levels of MIP-2. This stage could be reproduced by intradermal injection of IL-1alpha or IL-1beta and was partially blocked by injection of neutralizing Ab against IL-1alpha but not IL-1beta. In animals depleted of circulating neutrophils, chemokine expression was reduced by nearly 70% during this stage. In the second stage, which peaked at 24 h after injury, modest but significant levels of IL-1beta were detected in association with low levels of KC and high levels of MIP-2. This pattern of chemokine expression could not be mimicked by injection of IL-1alpha or IL-1beta (even with prolonged exposure), although MIP-2 expression could be partially inhibited by intradermal injection of neutralizing Ab against IL-1beta. Surprisingly, neutrophil depletion before injury resulted in sustained high levels of both KC and MIP-2 expression. These observations demonstrate that these two closely related chemokines are under distinct regulatory controls in vivo that are likely to reflect the temporally ordered participation of different cell types and/or extracellular stimuli and inhibitors.

Abdomen↗
PubMed1985

UV suppression of mast cell-mediated wealing in human skin.

Prior exposure of normal skin to subthreshold doses of UVR from a "UVA" phototherapy cabinet resulted in a small but significant reduction of the wealing response to intradermally injected codeine phosphate, a mast cell degranulating agent. The flare component of the reaction was not affected, nor were the responses to intradermally injected histamine. Both UVA and UVB from banks of filtered fluorescent tubes appeared to produce a similar but less pronounced effect. Dose-response studies revealed that the inhibition of mast cell-mediated wealing required a certain cumulative threshold dose of UVR (about 35 J/cm2 UVA + 20 mJ/cm2 UVB). The observed reduction in wealing could not be entirely explained by differences in the content of tissue histamine. It is concluded that repeated exposure to a broad range of UV wavelengths can depress the response of mast cells in vivo to degranulating agents by an unknown mechanism. This finding may have relevance to some of the observed therapeutic effects of phototherapy in certain inflammatory dermatoses and especially cutaneous mast cell disorders.

Adult↗
PubMed1996

Peripheral blood eosinophilia in owl monkeys is associated with increased eosinophilopoiesis yet depressed recruitment kinetics to the Chemokine RANTES.

New world nonhuman primates of the genus Aotus (owl monkeys) can be categorized by 11 distinct karyotypes (K). It has been demonstrated that monkeys of K-VI persistently have one order of magnitude more eosinophils (EOS) in the peripheral blood than K-I monkeys. The purpose of this study was to investigate the basis for this difference and examine EOS recruitment using two cutaneous models of inflammation. Peripheral blood EOS were isolated on metrizamide gradients to > or = 95% purity and then used for phenotypic studies. There were no significant differences when comparing karyotypes in the ratio of normodense (K-I, 6.4% +/- 3.8%; K-VI, 21.1% +/- 8.8%) EOS or their survival in culture (K-I, 5.3% +/- 2.9% at 72 hours; K-VI, 2.8% +/- 0.7% at 72 hours) (P > .05). Examination of bone marrow revealed that K-VI monkeys had greater than fivefold more EOS and EOS precursors than K-I animals. To examine EOS function in recruitment, monkeys of each karyotype were given a single intradermal injection of Escherichia coli lipopolysaccharide (LPS) or human recombinant (PMN) and mononuclear cells occurred in response to LPS as early as 4 hours after injection; the severity of infiltration was similar in both karyotypes and at all time points up to 24 hours. In contrast, by 8 hours after intradermal injection of RANTES, leukocyte infiltration in K-I monkeys consisted mostly of PMN (94.8% +/- 0.7%) that were predominantly EOS. In comparison, there was essentially no infiltrate in K-VI animals at all time points. There was no difference in VCAM-1 expression in response to intradermal LPS or RANTES between the two karyotypes. These results suggest that the genetic basis of peripheralblood eosinophilia in K-VI owl monkeys is likely a function of heightened eosinophilopoiesis and depressed recruitment kinetics from the peripheral circulatory pool in response to RANTES.

Animals↗
PubMed1981

Vascular permeability activity of botulinum C2 toxin elicited by cooperation of two dissimilar protein components.

Botulinum C2 toxin has vascular permeability as well as lethal activities. Both activities are elicited by cooperation of two dissimilar protein components, designated components I and II, which individually have very low activities. The vascular permeability activity of C2 toxin, demonstrated as blueing response after intravenous injection of Evans blue, was markedly enhanced by treatment with trypsin and was abolished by neutralization with either anti-component I or II serum. Inflammatory reactions, such as edema, congestion, and hemorrhage, were found at the site of intradermal injection of trypsinized C2 toxin. No vascular permeability activity was demonstrated by the intradermal injection of the toxin of Clostridium botulinum types A through F. These results indicate that C2 toxin has a novel biological activity, which is not possessed by the neurotoxin elaborated by C. botulinum types A through F. This suggests that C2 toxin causes lethality in a different way from that of botulinum neurotoxin, which is known to inhibit the presynaptic release of acetylcholine at the neuromuscular junction.

Animals↗
PubMed1989

Technetium-99m haemaccel: a new lymphoscintigraphic agent.

Haemaccel, a denatured gelatin polymer with average molecular weight of 35,000, was labeled with Tc-99m and evaluated in rats and rabbits. The labeling efficiency was consistently higher than 85%. Tc-99m haemaccel (200 microCi in 0.05 ml) was injected intradermally into the hind footpads of rats lightly anaesthetized with diethyl ether vapour. Using a gamma camera, static images of the anterior view were obtained at various time intervals after injection. The images obtained demonstrated good localization of Tc-99m haemaccel in regional lymph nodes. In rats, about 3% of the injected dose was trapped in the node, and 48% remained at the injection site at 4 h after injection. In rabbits, good-quality images of lymph nodes and lymphatic channels of the hind legs were obtained within 15 min after intradermal injection of Tc-99m haemaccel. Tc-99m haemaccel showed fast migration from the injection site, good accumulation in the lymph nodes, and good visualization of lymphatic channels. Hence, it has a potential application in lymphoscintigraphy.

Animals↗