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The purpose of this preliminary study was to determine the feasibility of ultrasound-guided fine-needle aspiration biopsy of suspected neoplastic lesions of bone. Ultrasound-guided fine-needle aspiration biopsy samples were obtained in 23 patients (22 dogs and one cat) with radiographic evidence of a destructive or destructive/productive bone lesion. The lesions were located in the appendicular skeleton in 20 patients and in the axial skeleton in three. Histopathology from tissue core biopsies and/or necropsy was not deemed necessary in 11 patients where ultrasound-guided fine-needle aspiration biopsy results were conclusive for neoplasia. A cytologic diagnosis from ultrasound-guided fine-needle aspiration biopsy was confirmed by histologic samples obtained at surgery or necropsy in five patients. In one of these five, ultrasound-guided fine-needle aspiration biopsy samples were diagnostic for sarcoma when tissue-core biopsy was inconclusive. Both ultrasound-guided fine-needle aspiration biopsy and tissue core biopsy techniques were inconclusive in one patient. Ultrasound-guided fine-needle aspiration biopsy samples were nondiagnostic in five patients where a follow-up histopathologic diagnosis of neoplasia was made. It was concluded that ultrasound-guided fine-needle aspiration biopsy of bone, if diagnostic, may help avoid the need for a tissue-core biopsy in some instances. However, a negative ultrasound-guided fine-needle aspiration biopsy sample does not rule out neoplasia. A negative ultrasound-guided fine-needle aspiration biopsy should be followed by a tissue-core biopsy and histologic analysis in all patients. Ultrasound-guided fine-needle aspiration biopsy seems to be a promising technique for the diagnosis of bone lesions.

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Routine serum protein electrophoresis is recognized as the most reliable assessment of avian protein profiles in health and disease and has replaced biochemical determination of albumin and A:G ratio in the ability to predict abnormalities of clinical significance. The importance of considerable species differences to the overall interpretation of avian electrophoresis is well established and constitutes a continued challenge to the avian specialist and to the providing laboratories to continue the pursuit of species-specific, even age- and gender-specific, reference ranges. Patterns for various diseases continue to emerge as more scrutiny is applied to the use of this tool in avian diagnostics for overall health assessment as an adjunct to specific disease diagnosis and for both prognostic and therapeutic monitoring.

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The reliability of utilizing cytologic samples for immunophenotyping canine lymphomas was evaluated by a systematic comparison of results from fine needle aspirate (FNA) to needle biopsy specimens stained by immunochemical methods. The specific reactivity of a selected panel of 12 antibodies to cell surface markers and intermediate filaments was assessed in 11 dogs by comparing cytologic to histologic samples. There was excellent correlation of results between immunostained cytologic and histologic samples. FNA sampling is a simple, noninvasive method for determining the lymphoid phenotype in canine lymphoma. In addition, this technique may be useful as a cytodiagnostic aid in differentiating lymphoid from non-lymphoid tumors and in assessing neoplastic vs. reactive or hyperplastic processes.

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It has been recently demonstrated that two serotypes of type D simian retroviruses, namely SRV-1 and SRV-2, exhibit extensive immunological cross-reactivity but do not exhibit cross-reactivity at the level of neutralizing antibodies. We have also shown recently that an area which includes residues 147-162 of the envelope protein of SRV-1 constitutes an epitope to which neutralizing antibodies against SRV-1 but not against SRV-2 are directed. However, in spite of the capacity of various immunogenic preparations to induce antibodies which react with SRV-1 these antibodies were incapable of neutralizing in vitro viral infectivity. Work reported herein demonstrates that various immunogens consisting of a larger peptide, namely 142-167 of the envelope protein of SRV-1, induce antibodies capable of binding with the envelope protein of SRV-1 and with the whole virus. Moreover, these antibodies exhibit the capacity to neutralize in vitro the infectivity of SRV-1 but not of SRV-2.

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We previously reported that an area encompassing amino acids 147-162 of the envelope region of the simian (type D) retrovirus serotype 1 (SRV-1) constitutes an antigenic site for the binding of murine and rhesus neutralizing antibodies. Neutralizing antibodies to SRV-2 are directed to a different area, encompassing residues 96-102 of SRV-2. This paper presents data on the activity of an SRV-1 recombinant envelope protein (rEP) and of monoclonal hybridoma cell line, C11B8, produced from murine spleen cells immunized with SRV-1 rEP. Purified monoclonal antibodies from C11B8 bind to the SRV-1 rEP and to both SRV-1 and SRV-2. However, the monoclonal antibody exhibits strain specificity in the capacity to neutralize SRV-1 infection in vitro. Thus, C11B8 neutralizes SRV-1 infection but fails to neutralize four other known serotypes of the virus. C11B8 also binds to an SRV-1 synthetic peptide representing residues 142-167, which encompasses the previously defined antigenic site of recognition for neutralizing antibodies to SRV-1. This paper also contains evidence that the SRV-1 rEP construct binds the site for SRV-1 attachment to the cell receptor. This is indicated by the ability of SRV-1 rEP to compete with SRV-1 (but not with SRV-2) and inhibit its infectivity in vitro. In addition, SRV-1 rEP inhibits the neutralizing activity of C11B8 against SRV-1 infection in vitro. SRV-1 rEP has no inhibitory effect on rhesus neutralizing antibodies to SRV-2. Taken together, the above findings indicate that immunity conferred at the level of neutralizing antibodies during SRV infection is strain-specific and involves the recognition of envelope sequences unique to each strain.

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We previously demonstrated that the area encompassing residues 147-162 of the envelope protein of simian retrovirus serotype-1 (SRV-1) is the target for rhesus anti-SRV-1 neutralizing antibodies. A peptide representing amino acids 142-167 of envelope protein (gp70) is capable of inducing virus-neutralizing antibodies in mice. The virus receptor was immunoprecipitated from Raji cells using gp70 as the ligand. Antibodies to peptide 142-167 inhibit the immunoprecipitation, indicating that the receptor recognizes residues 142-167 of the viral envelope protein.

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An area encompassing residues 142-167 of the envelope protein of type D simian retrovirus (SRV-1) has been shown to contain the epitope to which neutralizing antibodies are directed. This area has been synthesized and shown to bind to monkey and mouse antiviral antibodies and to a virus neutralizing mouse monoclonal antibody. Protein conjugates of this peptide as well as the cross-linked or the free peptide induce antibodies capable of neutralizing, in vitro, viral infectivity. The cell receptor to the virus was isolated following extraction of Raji cells with non-ionic detergents. The receptor was isolated and characterized following radioimmuno-precipitation of 125I labeled cell extract bound to viral envelope protein. This immunoprecipitation could be inhibited by antiserum to peptide 142-167. Analysis in gels indicate that the receptor is of molecular weight of approximately 60 KDa. These results indicate that the neutralizing antibodies and the receptor recognize the same area on the viral envelope protein and that neutralization is the result of blocking the virus-receptor interaction by antibodies.

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There are at least three distinct serotypes of simian type D retrovirus (SRV) which exhibit extensive serological cross-reactivity, but no cross-reactivity exists at the level of serum neutralizing antibodies. Amino acid sequence analysis and hydrophobicity plots of SRV-1 and SRV-2 envelope proteins were compared in order to identify unique potential antigenic determinants to which respective neutralizing antibodies may be directed. Peptides representing residues 147-162 of SRV-1 and 96-102 of SRV-2 were synthesized and assessed for their immunoreactivity. Free peptide inhibition of strain-specific serum (rhesus) neutralizing antibodies to SRV-1 and SRV-2 was demonstrated using the SRV-1 147-162 peptide and the SRV-2 peptide, 96-102, respectively. Inhibition of serum neutralizing activity by these peptides was also strain-specific, showing no cross-inhibition. SRV-1 147-162 conjugated to a protein carrier and cross-linked to Sepharose beads specifically adsorbed neutralizing antibodies from SRV-1 immune rhesus sera. The antibodies eluted from the immunoadsorbent possessed SRV-1 neutralizing activity, but showed no effect on the infectivity of SRV-2. Peptide SRV-1 147-162 also exhibited the capacity to bind specifically with a mouse monoclonal antibody which neutralizes the infectivity of SRV-1. Mice immunized with a recombinant SRV-1 envelope protein or with whole, inactivated SRV-1 produced antibodies which bound the SRV-1 147-162 conjugate, but not the protein carrier itself. Mouse antibodies to the SRV-1 147-162 conjugate exhibited specific binding with both native SRV-1 and with recombinant SRV-1 envelope protein. These findings provide strong evidence that SRV-1 147-162 and SRV-2 96-102 constitute at least two unique antigenic determinants, or parts thereof, which participate in the strain-specific neutralizing antibody response. Moreover, the findings indicate that the SRV-1 neutralizing antibodies produced by monkeys and at least a certain population of neutralizing antibodies produced by mice recognize the same epitope of SRV-1.

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Rheumatoid factor (RF) was found in titers greater than 1:8 in 72.2% of cases of classical or definite canine rheumatoid arthritis (RA) and in 5.9% or normal sera. Serum fractionation and immunoabsorbant studies that much of the RF present was IgG, although activity was demonstrated in all 3 major immunoglobulin classes. Evidence of involvement of both IgG and IgM to form complexes of varying sizes was obtained.

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Laboratory studies were performed on 3 Collies with familial canine dermatomyositis, 6 progeny from a breeding of 2 of the Collies (incross litter), and 4 progeny from the breeding of an affected Collie male and a normal Labrador Retriever (outcross litter). Hematologic abnormalities were leukogram changes consistent with inflammation, and, in 2 severely affected incross dogs, anemia of inflammatory disease. Serum muscle enzyme activities were not markedly increased. High concentrations of immune complexes and proportionally increased total immunoglobulin G were present in the sera of moderately and severely affected incross dogs. The same dogs had weakly positive direct Coombs' tests, and 1 affected outcross dog had a strongly positive direct Coombs' test. Rheumatoid factor was present in a severely affected incross dog. A few dogs had polyclonal globulin increases that were attributed to inflammation. Low antibody titers to canine calicivirus were present in 3 dogs and a low titer to canine coronavirus was present in 1 dog.

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Treatment with sulfadiazine-trimethoprim caused serious, but reversible, allergic drug reactions in 6 Doberman Pinschers 10 to 21 days after the first drug exposure and/or within 1 hour to 10 days after reexposure. Nonseptic polyarthritis was found in all dogs. Glomerulonephropathy, focal retinitis, polymyositis, skin rash, fever, anemia, leukopenia, and thrombocytopenia were found in some dogs. These clinical abnormalities were typical of an immune-mediated vasculitis and mimicked other immune-mediated disorders. In a drug challenge study, 1 dog was given sulfadiazine and trimethoprim separately. Administration of trimethoprim alone did not result in any abnormalities; however, exposure to sulfadiazine caused recurrence of the polyarthritis, glomerulonephropathy, and focal retinitis within 5 days, suggesting that sulfadiazine likely was the offending agent in all cases. In addition, during the sulfadiazine reexposure, marked complement activation was documented at the time clinical signs were apparent, supporting the suggestion that sulfadiazine caused an immune complex disease (type-III hypersensitivity reaction). Since all dogs were of the same breed, a genetic predisposition of some Doberman Pinschers to react adversely to sulfadiazine was suspected.

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A dog with polyarthritis, angular joint deformities, and a high serum antinucleolar antibody titer was treated over a period of 20 months. During the clinical course, leukopenia, thrombocytopenia, Coombs' positive hemolytic anemia, and a pemphigus-type skin disorder developed, all of which responded to immunosuppressive therapy. It is not known whether the polysystemic disease in this dog represents a pleomorphic manifestation of canine systemic lupus erythematosus or multiple autoimmune disorders occurring in the same animal.

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Direct Coombs' antiglobulin tests were performed on 80 dogs with patent Dirofilaria immitis infection and 170 dogs negative for microfilaria of D. immitis. Presence or absence of anemia was determined by hematocrit in 55 of the heartworm negative dogs and 68 of the dogs with heartworm disease. Heartworm infected dogs showed a higher incidence (37%) of anemia than noninfected dogs (14.5%). Anemia was most prevalent in two groups of dogs with heartworm infection, one group showing vena caval syndrome (91%) and the other occult dirofilariasis (62.5%). These latter two groups of dogs also showed a significantly higher number of positive Coombs' reactions at 37 degrees C than other dogs with heartworm disease and the noninfected dogs. The number of positive Coombs' reactions at 4 degrees C among the total of 80 dogs with heartworm infection was significantly higher than that for dogs without heartworm disease. However, there was no positive correlation between anemia and the outcome of the Coombs' test at either temperature. These findings do not suggest that immunologic factors play a major role in the pathogenesis of anemia in dogs with heartworm disease.

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Immune-mediated disorders in cats share many clinical and pathologic similarities with their counterparts in other species. Cats, however, are unique among domestic animals owing to the involvement of feline leukemia virus. In addition, a number of other infectious organisms can produce immune-mediated sequelae--that is, FIP virus, FeSFV, and H. felis. Therefore, the diagnostic and therapeutic aims in the management of feline immune-mediated disorders must take into account the probability of a primary or underlying disease process.

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Histopathologic, immunopathologic and clinical evaluation of two hundred thirty dogs resulted in a diagnosis of canine autoimmune skin disease (AISD) in eighty four of the suspected cases. Pemphigus foliaceus was the most common diagnosis, followed by bullous pemphigoid and pemphigus vulgaris. The highest percentage of cases were mixed breeds, followed by shelties, collies, German shepherds and poodles. There was no apparent age or sex predilection. Eighty seven percent of the eighty four cases of AISD were diagnosed on the basis of supportive clinical criteria in addition to compatible histopathologic and/or direct immunofluorescent findings. Thirteen percent of the cases with a final clinical diagnosis of AISD lacked supportive immunohistopathologic criteria on biopsy and the diagnosis was based upon clinical appearance, exclusion of other dermatoses, and response to immunosuppressive therapy. This latter group included four cases of SLE diagnosed using a combination of clinical criteria and a positive test for antinuclear antibody. Thirty three percent of the AISD cases were diagnosed on the basis of compatible findings on both histopathology and direct immunofluorescence. A histopathologic diagnosis of AISD was made in sixty nine percent of the cases. Positive direct immunofluorescence was found in fifty two percent of the cases. Thirty five percent of the AISD cases were diagnosed on histopathologic criteria without supportive immunofluorescent findings, while nineteen percent had positive direct immunofluorescence with nonspecific histopathologic changes. The number of definitive positive results was greater for direct immunofluorescence (38%) than for histopathology (25%). There was an equal incidence (21%) of false positive results among non-AISD cases with both tests. Indirect immunofluorescence appears to have little value in the diagnosis of canine AISD. The recommended diagnostic approach to canine AISD involves the use of both histopathologic and immunopathologic evaluations.

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