Molecular Cloning and Functional Characterization of Human MIP-1δ, a New C–C Chemokine Related to Mouse CCF-18 and C10

We have isolated a novel human C–C chemokine, MIP-1, from a human fetal spleen cDNA library. The human MIP-1 cDNA has an unusually long 400-bp 5-prime untranslated region and a predicted 113-amino acid protein of 10 kDa. The coding sequence contains a signal peptide of 21 amino acids, indicating that the mature protein has 92 amino acids (8 kDa). Recombinant human MIP-1 produced by transfected human embryonic kidney 293 cells produced an 8-kDa protein, which confirmed the presence of a signal peptide. Compared with other human C–C chemokines, human MIP-1 shows the highest homology with human HCC-1, CK-8, murine C10, and CCF18 (MIP-1). The human MIP-1 gene is localized on chromosome 17 where most of the C–C chemokine superfamily is located. Human MIP-1 is expressed in T and B lymphocytes, NK cells, monocytes, and monocyte-derived dendritic cells, but not in bone marrow-derived dendritic cells. Its expression can be induced by other proinflammatory cytokines in monocytes and dendritic cells. Human MIP-1 is chemotactic for T cells and monocytes, but not for neutrophils, eosinophils, or B cells. Human MIP-1 induced calcium flux in human CCR1-transfected cells.     

Spontaneous and agonist-induced openings of an acetylcholine receptor channel composed of bovine muscle α-, β and δ-subunits

During the development of mammalian muscle the -subunit of the nicotinic acetylcholine receptor (AChR) is replaced by the -subunit to produce well-defined alterations in the conductance and gating of the channel. To gain a better unterstanding of the functional role of the and -subunits, we have studied the properties of an AChR channel lacking these subunits. The AChR expressed in Xenopus oocytes injected with the bovine -, and -subunit-specific mRNAs (referred to as -AChR) is unusual in that its channel opens spontaneously at a high frequency in the absence of agonist. From a comparison of the -AChR with complete receptors containing either the or -subunit, we conclude that the and -subunits influence most channel properties, including agonist binding, and are especially important for stabilizing the closed state of the unliganded receptor channel. The -AChR can form when a complete set of four subunit-specific mRNAs is injected. The ease with which it is assembled raises the possibility that the - AChR contributes to some of the variations in receptor properties that occur during development.     

γδ T Lymphocytosis Associated with Common Variable Immunodeficiency1

We present the case of a 28-year-old Caucasian female with common variable immunodeficiency (CVID) since age 5 who had a long history of hospitalizations for unexplained fevers and pulmonary infiltrates. The patient developed mild lymphocytosis 7 months prior to our evaluation. Flow cytometry of peripheral blood revealed an expansion of T lymphocytes, mild CD4 T lymphocytopenia, and a reduced CD4/CD8 ratio (0.2). Two subpopulations of T lymphocytes were found (CD3⁺/CD4^–/CD8⁺, 47%; CD3⁺/CD4^–/CD8^–, 53%), the vast majority of which expressed V-1. An infectious cause for the patient's T lymphocytosis could not be found. The sputum was chronically colonized with Staphylococcus aureus, and the organism produced TSST-1 in vitro. A bronchoalveolar lavage (BAL) revealed marked lymphocytosis, but T lymphocytes were not overrepresented in the BAL. Lymphocyte functional studies revealed poor proliferative responses to mitogens and staphylococcal superantigens and diminished cytokine production. V-1 T lymphocytes from the patient's blood were not expanded in vitro in response to staphylococcal superantigens. TCR gene rearrangement studies confirmed the presence of J and J1 clonal rearrangements accounting for only a small subpopulation of the T lymphocytes. These studies were repeated 5 months later and were unchanged. A bone marrow biopsy was negative for leukemia. Hence, the cause of the patient's T lymphocytosis could not be determined despite evaluation for underlying malignancy, occult infection, or superantigen-driven stimulation. The patient ultimately died of progressive respiratory insufficiency. The state of current knowledge regarding T lymphocytosis, decreased production of T lymphocytes, and a low CD4/CD8 ratio in association with CVID is discussed.     

Analysis of γδ+ T cells in peripheral blood of children with perinatal human immunodeficiency virus (HIV) infection

The present study examined CD8 antigen expression and variable (V) gene segment usage by T cell receptor (TCR)-⁺ lymphocytes in peripheral blood of symptomatic children with perinatal HIV infection. The relative number of ⁺, CD8⁺ T cells in most of the infected children was higher than that in uninfected children from HIV⁺ or HIV^– mothers and correlated with the immunodeficiency status of the patients. Infected infants and children over 1 year old also showed an increased proportion of V1-J1⁺ T lymphocytes. CD8 expression on those cells was higher in infected than in uninfected infants and children. Sequence analysis of the gene rearrangement of the predominant V1 family in peripheral blood of three HIV⁺ donors revealed extensive junctional diversity. These results suggest that the V skewing in the majority of HIV⁺ children reflects peripheral expansion of V1-J1⁺ T lymphocytes early in life, which might be involved in the mechanisms of HIV-induced immunodeficiency.     

The role ofγδ T cells in the normal and disordered immune system

Summary A small population of T cells does not express the conventional T cell receptor characterized by the and polypeptide chains (TCR) but instead, two polypeptides termed and (TCR). This alternative receptor is able to recognize antigen. It appears early in T cell ontogeny, but its role in the thymus prior to the availability of TCR remains unclear. In selected sites such as skin or gut TCR predominates in mice which might suggest a role of T cells in the first line of defense against infection, T cells secrete lymphokines and display cytotoxic activity. However, their activation requirements may differ from what is known for T cells since MHC-nonrestricted and also CD4 and CD8 negative T cells have been described. Preferential activation by mycobacterial antigens possibly indicates a special repertoire of the T cells. In various diseases slightly increased numbers of T cells were found, but these preliminary studies have not yet provided evidence for a major pathogenetic role of T cells.List of abbreviations C constant region (immunoglobulin or TCR gene segment) - CD4 cluster of differentiation 4 (mainly on helper cells) - CD8 cluster of differentiation 8 (mainly on cytotoxic cells) - D diversity region (immunoglobulin or TCR gene segment) - DNA desoxyribonucleic acid - IL2 interleukin 2 - J joining region (immunoglobulin or TCR gene segment) - kD kiloDalton - MHC major histocompatibility complex - NK natural killer (cells) - RA rheumatoid arthritis - TCR T cell receptor - V variable region (immunoglobulin or TCR gene segment)     

δ-opioid receptor antagonists exhibit properties of partial δ-receptor agonists in isolated perfused heart

Perfusion of the isolated intact rat heart with Krebs—Henseleit solution containing agonists ((-)-TAN-67, DPDPE, and dalargin) or antagonists of -opioid receptors (naltrindole, TIPP[], and ICI 174,864) in a final concentration of 0.1 mg/liter was followed by a decrease in the heart rate, end-diastolic pressure, contraction rate, relaxation rate, and left ventricular developed pressure. Perfusion with a solution containing the -opioid receptor agonist DPDPE or -antagonists naltrindole, TIPP[], and ICI 174,864 before modeling of global ischemia increased the severity of reperfusion-induced contractile dysfunction in the myocardium. Our results suggest that -opioid receptor antagonists in vitro exhibit properties of partial - receptor agonists.Translated from Byulleten Eksperimentalnoi Biologii i Meditsiny, Vol. 138, No. 10, pp. 424–427, October, 2004     

Far-field somatosensory evoked potentials in response to selective stimulation of small diameter myelinated fibers in the cat

Summary Five far-field components were identified preceding the cortical wave in response to the stimulation of the A fibers of the sural nerve. When A activity was also included in the afferent volley, a second cortical wave and additional far-field potentials were recorded. Upon blocking the A fiber activity and thus isolating the peripheral input to A fibers alone, the initial cortical wave and the far-field potentials preceding it disappeared completely, leaving the A evoked potentials intact.     

δ-opioid receptor antagonists exhibit properties of partial δ-receptor agonists in isolated perfused heart

Perfusion of the isolated intact rat heart with Krebs—Henseleit solution containing agonists ((-)-TAN-67, DPDPE, and dalargin) or antagonists of -opioid receptors (naltrindole, TIPP[], and ICI 174,864) in a final concentration of 0.1 mg/liter was followed by a decrease in the heart rate, end-diastolic pressure, contraction rate, relaxation rate, and left ventricular developed pressure. Perfusion with a solution containing the -opioid receptor agonist DPDPE or -antagonists naltrindole, TIPP[], and ICI 174,864 before modeling of global ischemia increased the severity of reperfusion-induced contractile dysfunction in the myocardium. Our results suggest that -opioid receptor antagonists in vitro exhibit properties of partial - receptor agonists.Translated from Byulleten Eksperimentalnoi Biologii i Meditsiny, Vol. 138, No. 10, pp. 424–427, October, 2004     

Morphologische Untersuchungen an pathogenen und potentiell pathogenen Amoeben der Typen „Entamoeba“ und „Hartmannella-Acanthamoeba“ aus Reptilien

Zusammenfassung Es wurden 114 Stämme von reptilpathogenen und potentiell reptilpathogenen Amoeben vom Typ Hartmannella-Acanthamoeba und der Gattung Entamoeba mit Hilfe der Single Linkage Cluster Analysis (SLCA) auf Grund von morphologischen Merkmalen in fünf Typen und einige Außenseiter aufgegliedert.Dabei zeigte sich, daß die Hartmannellen eine einzige Gruppe bilden, die einen sehr hohen Homogenitätsgrad besitzt. Innerhalb der heterogenen Gattung Entamoeba konnten 4 Typen (1, 2, 3 und 4) definiert werden, von denen Typ 1 und 2 Varianten der gleichen Art sein dürften, während Typ 3 und 4 wahrscheinlich unterschiedliche Arten sind. Der Unterschied zwischen den Untergruppen (1 und 2 und 3 und 4) ist größer als der zwischen den Typen 3 und 4.Eine eindeutige Zuordnung bestimmter Amoebentypen zu bestimmten Reptilgattungen war nicht möglich. Auch ließen sich krankheitserregende Formen nicht von solchen unterscheiden, die nur zu latenten Infektionen führten. Alle Typen konnten als pathogene oder auch als latente Erreger auftreten.

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Morphological study of pathogenic and potentially pathogenic amoebae of the types Entamoeba and Hartmannella-Acanthamoeba from reptiles

Summary A total of 114 strains of reptile pathogenic or potentially pathogenic amoebae of the genus Hartmannella-Acanthamoeba and Entamoeba was partitioned into five types and several outsiders by means of Single Linkage Cluster Analysis (SLCA) based on morphological items. The Hartmannella strains proved to be fairly homogenious whereas within the heterogenious group of Entamoeba strains four types (no. 1, 2, 3, and 4) could be defined.Types 1 and 2 are likely to be varieties of the same species whereas types 3 and 4 are probably distinct species. Difference between subgroups (1, 2) and (3, 4) is greater than that between types 3 and 4. A distinct association of specific amoebae types to specific reptile orders failed. Furthermore, pathogenic forms could not be distinguished from those causing only latent infections. Heavy amoebiasis and latent infections as well were due to all four types.

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Die Arbeit wurde mit Unterstützung der Deutschen Forschungsgemeinschaft durchgeführt.     

δ Sequences mediate DNA rearrangements in Saccharomyces cerevisiae

Summary A solo sequence flanking the 5 end of the ADHII structural gene, ADR2, can promote a number of DNA rearrangements some of which were investigated in detail. In a selective system haploid mutants were screened in which a solo S sequence flanking ADR2 had been joined to a Ty element. Three different types of events can create such a structure: Reintegration of a Ty sequence at the -ADR2 site, inversion of ADR2 and flanking material, and transposition of ADR2 along with 3 flanking material. The involvement of reciprocal or non-reciprocal exchange mechanisms in creating such events are discussed.     

Increase in TCRγδ T lymphocytes in synovia from rheumatoid arthritis patients with active synovitis

To determine the relative presence of TCR+ and TCR+ T cells in synovial tissue from patients with various types of inflammatory synovitis and in tissues from patients with a number of chronic T cell-mediated conditions, we stained frozen tissue sections with monoclonal antibodies in indirect immunofluorescence assays. In tissues obtained from patients with chronic T cell-mediated granulomatous conditions (Wegener's granulomatosis, lymphomatoid granulomatosis, granuloma annulare, Langerhan's cells granulomatosis, pigmented villonodular synovitis, Takayasu's arteritis, and talc granulomatosis), the T cells present were predominantly TCR+, without an increased presence of TCR+ cells. In contrast, 6 of 14 (43%) synovia from patients with rheumatoid arthritis (RA) showed increased TCR+ T cells (3–10 cells/hpf). The RA synovia with increased TCR+ cells present had an increased tissue inflammation score compared to RA synovia with few TCR+ cells [18.6±5.8 versus 11.6±4.2 (mean±SE),P<0.05]. In contrast, synovia from patients with osteoarthritis, systemic lupus erythematosus, and trauma did not show an increased presence of TCR+ T cells. Thus, in cellular inflammatory infiltrates the presence of increased TCR cells is not a component of noninfectious granulomatous inflammation but is found in approximately 40% of RA synovia with high levels of inflammation.     

Insulin therapy prevents spontaneous recovery from streptozotocin-induced diabetes in Syrian hamsters

Summary Streptozotocin (Sz) given as a single dose of 50 mg/kg body wt. caused severe diabetes in Syrian hamsters. However, the level of blood glucose decreased gradually after 21 days post-Sz and reached the near normal level at 70 days in 90% of hamsters. The recovery from diabetes was associated with the regeneration of the-cells of islets and a reduction in the initially increased number of- and-cells. Daily treatment of diabetic hamsters with insulin was associated with the persistence of severe diabetes, lack of or minimal tendency for-cell regeneration and sustained hyperplasia of- and-cells in 90% of hamsters. Insulin also inhibited DNA synthesis (as measured by incorporation of tritiated thymidine), in ductal, ductular and acinar cells in Sz-pretreated hamsters but not in normoglycemic control hamsters treated with insulin alone. The results demonstrate a deleterious effect of exogenous insulin in the course of Sz-induced diabetes in hamsters.This work supported by grant CA38922, Laboratory Core grant CA36727, NCI/NIH, and SIG-16, American Cancer Society     

A T cell/B cell/epithelial cell internet for mucosal inflammation and immunity

Conclusions Mucosal immune responses are strongly regulated by CD4⁺ T cells and their derived cytokines. In this regard, IFN-^–/– mice (i.e., which lack Th1 and have elevated Th2 cells) showed strong mucosal Th2-type responses together with S-IgA production, while IL-4^–/– (e.g., dominant Th1 and lack of Th2 cells) mice had impaired mucosal Th2 and IgA responses following oral delivery of TT and CT. However, when rSalmonella or radenovirus were used for antigen delivery, significant levels of mucosal IgA responses were induced in both IFN-^–/^– and IL-4^–/^– mice. The choice of the antigen delivery system which leads to optimal Th and B cell interactions are important for the induction of effective IgA responses, even in situations where the immune system is compromised. It is clear that Th2-type cytokines are important in mucosal IgA responses; however, other cytokine combinations can compensate for mucosal immunity in situations in which Th2 cell responses are absent. Mucosally induced tolerance may be one approach to prevent several systemic immune disorders; however, the mechanism of this phenomenon still needs to be elucidated. Our recent findings have suggested that IFN- may play an important role in induction of systemic unresponsiveness since oral tolerance was not induced in IFN-^–/^– mice.Our studies as well as those of others indicated that at least two phases of a triad of cell interactions are important for the mucosal immune system. First, it has been shown that epithelial cell-produced IL-7 and SCF and T cell-derived IL-2 are essential activation and growth signals for intestinal T cells. Second, our studies with TCR knockout mice have suggested that mucosal T cells also play a critical role in the regulation of mucosal IgA responses. Thus, a mucosal internet among T cells, T cells, and IgA B cells appear critical for mucosal homeostasis and for regulation of specific mucosal immune responses.     

Elevated numbers of gamma-delta (γδ+) T lymphocytes in children with immune thrombocytopenic purpura

Immune thrombocytopenic purpura (ITP) in childhood is a heterogeneous clinical disorder characterized by immune-mediated platelet destruction. Although generally considered to involve autoreactive B lymphocytes which produce antiplatelet antibodies, there is increasing evidence that T lymphocytes also play an important role in this autoimmune process. We studied 11 children with acute ITP and 19 children with chronic ITP and observed elevated numbers of TCR+ T lymphocytes in several patients. In the three children with the highest elevations (TCR+/CD3+ percentage ranging from 37.8 to 48.1% at initial evaluation), the expanded cell population exclusively expressed the surface V2/V heterodimer and had enhanced,in vitro proliferation to mycobacterial extracts and IL-2. Analysis of the nucleotide sequences used by these TCR+ cells demonstrated a diverse set of VDDJC gene rearrangements, indicating polyclonal expansion of cells reminiscent of a superantigen response. There was a close correlation between the number of TCR+ T lymphocytes and the degree of thrombocytopenia in each patient. TCR78+ T lymphocytes may be important in the pathogenesis of immunemediated platelet destruction in some children with ITP.     

Effects of Β blockade and atropinisation on plasma catecholamine concentration during exercise

The changes in plasma catecholamine concentration (C) following -blockade (practolol, 15 mg) and atropinisation (Atropine, 1.8 mg) have been studied on 5 healthy male subjects during exercise on a motor driven treadmill.The results showed that for a given VO₂ and % VO_{2 max}, blockade was without effect on C (except in one athletic subject), but atropine produced a rise in C. In relation to Q, both drugs produced an increase in C, but for a given cardiac frequency (f _H ) C was higher with blockade, and lower with atropinisation than found in control experiments. The intra- and inter-subject variability of C in relation to f _H was resolved by considering the change in cardiac frequency calculated from baseline value obtained during walking at 6.44 km/h on the level, and expressed as a percentage of the maximal f _H attainable for given individuals under the different drug and control conditions (% f _H ).It was concluded that during short term exercise, the rise of C in relation to % f _H reflects both the myocardial sensitivity to vagal and blockade, and the circulatory vasoconstrictor control of blood vessels which is required to sustain increases in systemic and muscle blood flow.     

Distribution of opiate receptors within visual structures of the cat brain

Summary The distributions of , , and opiate receptors within visual regions in the cat cortex, thalamus and midbrain were determined by in vitro autoradiography. The overall distribution of receptors was examined using [³H]-etorphine, a ligand that nonselectively labels all types of opiate receptors. [³H]-[D-Ala², N-Me-Phe⁴, Gly(ol)⁵]-enkephalin (DAGO) was used to selectively label receptors, [³H]-[D-Pen^2,5]-enkephalin (DPDPE) for receptors, and [³H]-bremazocine for receptors. Each of the areas examined showed clear opiate receptor binding with [³H]-etorphine and a differential distribution of , , and receptors. Compared to other cortical regions, opiate binding in layers 3 and 4 of areas 17 and 18 was sparse. In the adjacent areas a more uniform distribution across layers was observed. The density of opiate receptors was greater in cortex than in subcortical structures, whereas the reverse was the case for receptors. Nevertheless, all three types of opiate receptors were found in the ventral and dorsal subdivisions of the lateral geniculate (LGN), the pulvinar complex, and the suprageniculate nucleus. In the midbrain, the superficial layers of the superior colliculus were heavily labelled with the , receptor ligand, and modestly with the ligand. Compared with other midbrain and diencephalic areas, binding was low in the superior colliculus. These results suggest that the diverse effects of opiates on visual perception are mediated by the unique distributions of opiate receptor types throughout the visual areas in the brain.     

In situ expression of β1, β3 and β4 integrin subunits in non-neoplastic endothelium and vascular tumours

Endothelial cells play an important role in adhesive interactions between circulating cells and extracellular matrix proteins. In vitro studies have shown that many of these processes are mediated by a superfamily of heterodimeric transmembrane glycoproteins called integrins. The distribution patterns of 1, 3 and 4 integrin subunits in endothelial cells (EC) in situ were examined immunohistochemically on serial forzen sections of a wide range of non-neoplastic tissues and of vascular tumours, both benign and malignant. Expression of the 1 subunit was a constitutive feature of EC. Among the 1-associated subunits, 5 and 6 were broadly distributed in EC, irrespective of vessel size and microenvironment. The 3 subunit displayed intermediate levels of expression with a slight preference for small vessel EC. Presence of 1 was confined to EC of capillaries and venules/small veins. Expression of 2 in EC was inconsistent. With rare exceptions, the 4 chain was absent in EC. The 3 and v subunits were expressed in most EC, though not always concomitantly. In contrast to the 1 chain, however, these integrin subunits were absent in EC of glomerular capillaries and were expressed variably in sinusoidal EC. The 4 chain was evenly present in the great majority of EC, except for those of large vessels. In vascular tumours, the patterns of 1 and 1 to 6 subunit expression generally corresponded to those found in their non-neoplastic counterparts. Expression of 3, v and 4 chains, however, decreased in neoplasia, especially in angiosarcomas. These data show that EC dispose of broad and at the same time differential repertoires of integrin subunits that presumably reflect vessel-type associated functional differences among these cells. In vascular tumours, the orthologous distribution patterns of 1 and 1 to 6 chains are conserved in most instances while the amounts of 3, v and 4 subunits expressed in EC tend to decrease in the course of malignant transformation.Dedicated to Prof. Dr. med. Dres. h.c. Wilhelm Doerr on the occasion of his 80th birthday     

Interferon β1a Treatment Modulates TH1 Expression in γδ + T Cells from Relapsing-Remitting Multiple Sclerosis Patients

A paradigm exists that multiple sclerosis is causally related to dysregulation of T_H1 inflammatory cytokines and T_H2 antiinflammatory cytokines. The cytokine source(s) that initiate the imbalances are unknown. In this study, , CD4, and CD8 T cell receptor-positive (TCR+) cells were isolated from the blood of 26 definitive relapsing-remitting multiple sclerosis patients prior to interferon -1a (IFN1a) therapy and following 8–10 weeks of this therapy. The bioactivities of interferon (IFN), interleukin 10 (IL10), and interleukin 12 (IL12) were determined. The concentrations of IFN, IL10, and IL12 from each cell type did not change significantly with IFN1a treatment. The IL10 secreted by TCR+ cells strongly correlated with the IL12 secreted by the same TCR+ cells, supporting the paradigm. Furthermore, IFN1a therapy decreased the TCR+ cell secretion of T_H1 cytokines after 8–10 weeks of therapy.     

DNA polymerase δ of Physarum polycephalum

DNA polymerase from the phylogenetically ancient slime mold Physarum polycephalum has been 380-fold enriched from amoebae. It was found to have the properties typical for this type of DNA polymerase from higher eukaryotes with regard to effectors, template-primer acceptance, co-purification with 3–5-exonuclease activity, as well as the effect of endogenous proliferating cell nuclear antigen (PCNA) from amoebae on the stimulation and processivity of DNA synthesis. An identified cDNA fragment shows 65.5% identical amino acides with DNA polymerase from Saccharomyces pombe. The molecular mass of the polymerase is 125 kDa while that of PCNA is 35 kDa. During size-exclusion chromatography, the highly purified polymerase eluted in the position of 125 kDa, suggesting that no other proteins were tightly complexed with the enzyme. The DNA polymerases from the (mononucleate) amoebae and from the (multinucleate) plasmodia of P. polycephalum have very similar properties in contrast to their differences in phenotype and their mode of nuclear division. The polymerase shows a higher degree of similarity than DNA polymerase , and especially the -like DNA polymerase, with the corresponding polymerases of higher eukaryotes. According to antibody staining, DNA polymerase is readily fragmented by proteases, even in the presence of inhibitor cocktails. Including freshly prepared cell lysates, proteolytic fragments are reproducible, the most abundant being 50 kDa in size. The DNA polymerase is recognized by the antisera against two peptides which have been derived by PCR-screening of plasmodial cDNA. One of the proteolytic splitting sites is located within an eight amino-acid stretch between the two antigenic sequences.     

Analgesic and antiinflammatory activity of constituents ofCannabis sativa L.

Two extacts ofCannabis sativa herb, one being cannabinoid-free (ethanol) and the other containing the cannabinoids (petroleum), were shown to inhibit PBQ-induced writhing in mouse when given orally and also to antagonize tetradecanoyl-phorbol acetate (TPA) -induced erythema of mouse skin when applied topically. With the exception of cannabinol (CBN) and ¹-tetrahydrocannabinol (¹-THC), the cannabinoids and olivetol (their biosynthetic precursor) demonstrated activity in the PBQ test exhibiting their maximal effect at doses of about 100g/kg. ¹-THC only became maximally effective in doses of 10 mg/kg. This higher dose corresponded to that which induced catalepsy and is indicative of a central action. CBN demonstrated little activity and even at doses in excess of 10 mg/kg could only produce a 40% inhibition of PBQ-induced writhing. Cannabidiol (CBD) was the most effective of the cannabinoids at doses of 100g/kg. Doses of cannabinoids that were effective in the analgesic test orally were used topically to antagonize TPA-induced erythema of skin. The fact that ¹-THC and CBN were the least effective in this test suggests a structural relationship between analgesic activity and antiin-flammatory activity among the cannabinoids related to their peripheral actions and separate from the central effects of ¹-THC.