Activation of innate anti-viral immune response genes in symptomatic benign prostatic hyperplasia.

Benign prostatic hyperplasia (BPH) is the most common urologic disease in men over age 50. Symptoms include acute urinary retention, urgency to urinate and nocturia. For patients with severe symptoms, surgical treatment is used to remove the affected tissue. Interestingly, the presence of histologic BPH does not always correlate with symptoms. The molecular basis of symptomatic BPH and how it differs from asymptomatic BPH is unknown. Investigation into the molecular players involved in symptomatic BPH will likely give insight into novel therapeutic, and potentially preventative, targets. We determined the expression of genes involved in the innate anti-viral immune response in tissues from patients undergoing surgery to alleviate the symptoms of BPH, and compared the results with prostate tissue with histologic BPH, but from patients with few urinary issues (asymptomatic BPH). We found that expression of complement factor I, apolipoprotein B mRNA-editing enzyme, catalytic polypeptide-like protein 3G, oligoadenylate synthetase 2 and interferon-induced tetratricopeptide 1, four genes whose protein products are involved in the innate anti-viral immune response, was significantly transcriptionally upregulated in symptomatic BPH. Additionally, we observe hypomethylation and concomitant expression of ancient retroviral-like sequences, the long interspersed nuclear element 1 retrotransposons, in symptomatic BPH when compared with normal prostate tissue. These findings merit further investigation into the anti-viral immune response in symptomatic BPH.

Differential induction of H-2K versus H-2D class I major histocompatibility antigens by recombinant gamma interferon. Lack of Kk augmentation in a leukemia virus-induced tumor is due to a cis-dominant effect.

T-T tumor hybrids were constructed between the AKR SL3 thymoma and an H-2-distinguishable thymoma cell line. Hybrids were stimulated with IFN-gamma to determine whether the differential augmentation of H-2D vs. H-2K class I antigen expression by AKR SL3 in response to IFN-gamma was due to effects cis or trans to the noninducible Kk gene. For each of a large number of hybrids tested, the expression of H-2Db, Kb, and Dk, but not Kk, was substantially enhanced by murine rIFN-gamma. These results suggested that the lack of induction of the Kk gene was due to an alteration cis to Kk rather than to the presence or absence of K region-specific, trans-acting negative or positive factors, respectively.

The generation and specificity of cytotoxic T cells raised against syngeneic tumor cells bearing AKR/Gross murine leukemia virus antigens.

Efforts were made to generate C57BL/6 cytotoxic effector cells to a syngeneic leukemia (E{male}G2) bearing AKR/Gross virus antigens. As we were unable to induce significant cytotoxic activity by immunization with up to 10(8) irradiated E{male}G2 cells, even when cells from such primed animals were subsequently restimulated with E{male}G2 cells in vitro, C57BL/6 mice were immunized with an aliogeneic, virus-producing AKR leukemic cell line (AKR SL3). Peritoneal exudate cells and, to a lesser degree, spleen cells from these mice showed significant lytic activity toward the immunizing allogeneic tumor but not toward E{male}G2. When spleen cells were harvested from animals {approximately equal to}10 d after injection of AKR SL3 and rechallenged in vitro with either E{male}G2 or AKR.H-2(b) SL1, another tumor that displays AKR/Gross virus antigens, then a vigorous cytotoxic response against E{male}G2 and AKR. H-2(b) SL1 was obtained. Effector cells generated by AKR SL3 priming followed by in vitro stimulation with E{male}G2 or AKR.H-2(b) SL1 lysed only cells of H-2(b) haplotype which were strongly positive for the display of serologically detectable AKR/Gross virus antigens. Thus, AKR SL3 cells were not lysed nor were EL4 cells (H-2(b); but only weakly positive for gp70). Cells not bearing the MuLV antigens tested for, such as P815 mastocytoma cells and spleen cell "blasts" from C57BL/6 and CBA (H-2(k)) mice, were also insusceptible to attack. The cytotoxic effector cells induced bore Thy 1.2 alloantigen and were of the Lyt 1+2+ phenotype. Collectively, these findings are consistent with the conclusion that the cytotoxic T cells raised against E{male}G2 are directed against AKR/Gross virus-associated antigens and are H-2 restricted. It will be of interest to determine the relevance of such effector cells to the known resistance of the C57BL/6 mouse to AKR/Gross virus-induced leukemia.

Antibody to the ligand of CD40, gp39, blocks the occurrence of the acute and chronic forms of graft-vs-host disease.

Chronic and acute graft-versus-host disease (cGVHD and aGVHD) result from donor cells responding to host disparate MHC alleles. In cGVHD (H-2d-->H-2bd), heightened polyclonal immunoglobulin production is due to the interaction of donor allospecific helper T cells (Th) and the host B cells. In vivo administration of antibody to the ligand for CD40, gp39, blocked cGVHD-induced serum anti-DNA autoantibodies, IgE production, spontaneous immunoglobulin production in vitro, and associated splenomegaly. Antibody production remained inhibited for extended periods of time after termination of anti-gp39 administration. Antiallogeneic CTL responses induced in a GVHD were also prevented by the in vivo administration of anti-gp39 as was the associated splenomegaly. These data suggest that CD40-gp39 interactions are critical in GVHD and that CD40-gp39 may be a valuable ligand-receptor pair for targeting immunotherapeutic agents to control GVHD.

Cytotoxic activities of monoclonal antibodies against the envelope proteins of murine leukemia virus.

Monoclonal antibodies against the envelope proteins [gp70 and p15(E)] of murine leukemia virus react with the cell surface of virus-infected cells. The specificity and potency of these antibodies exceed those observed with conventional polyvalent antisera. In cytotoxic assays, certain of the monoclonal anti-gp70 antibodies demonstrate 1000-fold differences in their titer on leukemic and normal thymus cells.

Characterization of a human ovarian carcinoma cell line (NIH:OVCAR-3) with androgen and estrogen receptors.

A cell line, NIH:OVCAR-3, has been established from the malignant ascites of a patient with progressive adenocarcinoma of the ovary after combination chemotherapy with cyclophosphamide, Adriamycin, and cisplatin. OVCAR-3 grows as a cobblestone-like monolayer with foci of multilayering, is tumorigenic in athymic mice, clones in agarose, and has an abnormal karyotype which includes a homogeneous staining region and a double minute chromosome. The cultured cells and xenografts contain cytoplasmic androgen- and estrogen-binding macromolecules with the specificity of the respective steroid hormone receptors. These components have sedimentation coefficients of 7 to 9S in low-salt sucrose-density gradients, have dissociation constants of 250 and 9.6 pM, and are present at concentrations of 30 and 28 fmol/mg cytosol protein characteristic of androgen and estrogen receptors, respectively. OVCAR-3 is resistant in vitro to clinically relevant concentrations of Adriamycin (5 X 10(-8) M), melphalan (5 X 10(-6) M), and cisplatin (5 X 10(-7) M) with survival compared to untreated controls of 43, 45, and 77%, respectively. Furthermore, there are multiple histological similarities between the patient's original tumor, the cell line, and the transplantable tumor. These data indicate that OVCAR-3 may be of use for investigations as to the significance of androgens and estrogens and the mechanisms of cytotoxic drug resistance in ovarian cancer.

Aldose reductase expression in human diabetic retina and retinal pigment epithelium.

The conversion of glucose to sorbitol by aldose reductase (AR) and its subsequent intracellular accumulation have been implicated in the pathogenesis of diabetic cataracts. There is also evidence linking AR activity with retinal capillary basement membrane thickening in galactosemic rats, suggesting a possible role in diabetic retinopathy. In this study, we explored one feature of this issue by examining diabetic and nondiabetic eyes for immunoreactive AR. AR was immunohistochemically undetectable in the retinal pigment epithelia (RPE) and neural retinas of nondiabetic human eyes. Weak, focal staining for AR was present unilaterally in the RPE of 1 of 11 diabetic patients without pathologic ocular findings and in 43% of diabetic patients with mild ocular findings. Retinal positivity was found (unilaterally) in only 2 of 19 individuals from either of these mildly affected groups. Fifty-five percent of patients with background retinopathy demonstrated AR positivity in the RPE, and half of these expressed AR in the RPE of both eyes. Of the individuals with proliferative diabetic retinopathy, 87.5% showed bilateral staining in the RPE. Retinal positivity was present in 36% of background retinopathy and 75% of proliferative retinopathy cases, demonstrating a positive correlation between AR expression and the severity of the disorder. In weakly staining retinas, only the ganglion cell bodies, nerve fibers, and Müller cells were positive, whereas in intensely staining cases, virtually the entire retina, except for the rod outer segments, was positive. Eyes from patients who had had diabetes less than or equal to 6 yr were negative for AR, but those from long- term-diabetic patients (14-45 yr) manifested positively.(ABSTRACT TRUNCATED AT 250 WORDS)

Detection of CX3CR1 single nucleotide polymorphism and expression on archived eyes with age-related macular degeneration.

There is a significant genetic component in age-related macular degeneration (AMD). CX3CR1, which encodes the fractalkine (chemokine, CX3CL1) receptor, has two single nucleotide polymorphisms (SNPs): V249I and T280M. These SNPs are correlated with other aged-related diseases such as atherosclerosis. We have reported an association of CX3CR1 SNP and AMD. In this study we examined CX3CR1 SNP frequencies and protein expression on archived sections of AMD and normal eyes. We microdissected non-retinal, peripheral retinal and macular cells from archived slides of eyes of AMD patients and normal subjects. CX3CR1 SNP typing was conducted by PCR and restriction fragment length polymorphism analysis. CX3CR1 transcripts from retinal cells were also measured using RT-PCR. CX3CR1 protein expression was evaluated using avidin-biotin complex immunohistochemistry. We successfully extracted DNA from 32/40 AMD cases and 2/2 normal eyes. Among the 32 AMD cases, 18 had neovascular AMD and 14 had non-neovascular AMD. The M280 allele was detected in 19/64 (32 cases x2) with a frequency of 29.7%, which was significantly higher as compared to the frequency in the normal population (11.2%). We detected CX3CR1 expression in the various retinal cells. CX3CR1 transcript and protein levels were diminished in the macular lesions. This study successfully analyzed CX3CR1 SNP and transcript expression in microdissected cells from archived paraffin fixed slides. Our data suggest that the M280 allele, a SNP resulting in aberrant CX3CR1 and CX3CL1 interaction, as well as lowered expression of macular CX3CR1, may contribute to the development of AMD.

Pneumocystis carinii pneumonia complicated by lymphadenopathy and pneumothorax.

A case of generalized Pneumocystis carinii infection presented as hilar and mediastinal lymphadenopathy and was complicated by spontaneous pneumothorax. Extrapulmonary P carinii infection in patients with acquired immunodeficiency syndrome is rare, and pneumothorax is even rarer. The purpose of this report is to call attention to these atypical features of P carinii infection in patients with the acquired immunodeficiency syndrome.

Neurotoxic effects of cisplatin therapy.

Swelling of the optic disc and loss of tendon reflexes developed in a 3 1/2-year-old girl after 11 high-dose courses of cisplatin (cis-dichlorodiammine platinum [II]) therapy for recurrent sacrococcygeal teratoma. Postmortem examination demonstrated optic disc swelling; the spinal cord showed loss of myelinated fibers and gliosis of the dorsal columns. To our knowledge this case is the first in which the neuropathologic changes associated with cisplatin therapy have been demonstrated by postmortem examination.

Cerebral ocular Whipple's disease: a 62-year odyssey from death to diagnosis.

A 47-year-old white man with dementia, supranuclear ophthalmoplegia, and myoclonic ocular and facial jerks died in 1931. The case report in 1936 by Ford and Walsh diagnosed encephalitis. In 1993, we made a clinical diagnosis of Whipple's disease on the basis of the 1936 publication. We restudied the pathologic material and found, in addition to extensive encephalitis, PAS-positive material in only the eye, brain, spinal cord, and pituitary. Electron microscopy demonstrated free and intracytoplasmic microorganisms in the eye and brain. We review the history of cerebral ocular Whipple's disease and the implications from this case, which occurred before the development of antibiotics.

Neonatal seizures and retardation in a girl with biochemical features of X-linked adrenoleukodystrophy: a possible new peroxisomal disease entity.

Neonatal hypotonia, seizures beginning at 5 days, and severe retardation were noted in a girl with normal karyotype and biochemical evidence of impaired adrenal function. Postmortem examination at 14 months revealed malformative and destructive lesions of central gray and white matter, atrophy of adrenal cortex with striated adrenocortical cells, hepatic fibrosis, and PAS-positive macrophages in several organs. Pathologically and clinically, this patient most closely approximated neonatal adrenoleukodystrophy (ALD) and differed strikingly from X-linked childhood ALD. In contrast, biochemical changes resembled the abnormalities observed in X-linked ALD and differed from those in the neonatal form. The very-long-chain fatty acid accumulation characteristic of both disorders was demonstrated, but unlike neonatal ALD, the levels or metabolism of plasmalogens, pipecolic acid, phytanic acid, and bile acid intermediates were normal, and peroxisomes in a liver biopsy specimen were present in normal number and appeared enlarged. While the case resembles the recently reported entity of peroxisomal acyl-CoA oxidase deficiency, assignment to this category was excluded by immunoblot studies on postmortem liver, which revealed normal amounts of this enzyme. Correlation of clinical, morphologic, and biochemical data suggests that this case is an example of a so-far undescribed entity, and reinforces the concept that the phenotypic spectrum of peroxisomal disorders is wider than realized.

Fatal viscerotropic Rocky Mountain spotted fever. Report of a case diagnosed by immunofluorescence.

A case of fatal viscerotropic Rocky Mountain spotted fever with virtual absence of cutaneous lesions was diagnosed at autopsy by specific immunofluorescent demonstration of Rickettsia rickettsii in spleen, kidney, epididymis and skin. The clinical presentation was that of insidious onset of fever, renal failure, hypotension, hyponatremia and obtundation over a 10 day period. The patient had respiratory insufficiency, hypocalcemia, increases in creatinine phosphokinase (CPK), serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), lactic dehydrogenase (LDH), alkaline phosphatase, billirubin and serum phosphate, grand mal seizure, myalgia and unremitting shock with death occurring on day 12 of illness. Postmortem examination revealed severe vasculitis with interstitial nephritis and multifocal tubular necrosis, pericholangitis with bile stasis, glial nodules in the brain, multifocal rhabdomyonecrosis, interstitial pneumonitis and mild interstitial myocarditis. Risk factors which this patient shared with other patients with fatal Rocky Mountain spotted fever were failure to recognize a rash, failure to obtain a tick bite history, male sex, black race and age greater than 30 years.

Cytomegalovirus retinitis in adults. A manifestation of disseminated viral infection.

Retinitis caused by cytomegalovirus (CMV) infection is unusual in adults. Sixteen of the 17 cases reported have occurred in immunologically compromised patients, most frequently renal transplant recipients. CMV retinitis is associated with a distinctive ophthalmoscopic appearance and, in the majority of cases, was the first clinical manifestation of systemic viral infection. Severe and permanent visual deficits are characteristic. Since retinitis is a reliable sign of disseminated disease and ophthalmoscopic examination a rapid method of establishing its presence, recognition of this manifestation should allow earlier diagnosis of serious CMV infection.

Retinal atrophy induced by intravitreous colchicine.

Colchicine is known to inhibit axoplasmic transport in ganglion cells. Previous studies have shown considerable, but largely reversible, retinal changes after low dosages of intravitreal colchicine in experimental animals. In the present study, the effects of 1.0 to 100 micrograms of intravitreal colchicine in monkeys were studied by ophthalmoscopy, light microscopy, and electron microscopy. Optic atrophy was ophthalmoscopically evident within 4 weeks after a single dose of 10 micrograms or more. Morphologic changes included progressive swelling of retinal neurons, accumulation of a fibrillogranular material, displacement of organelles, and loss of microtubules. Plasmalemmal rupture was observed in ganglion cells and in photoreceptors after as little as 1 microgram of colchicine. Relative sparing of the cone cells was noted. The retina of monkeys appears to be more sensitive to intravitreal colchicine than that of certain lower animals.

Ocular inflammation in autoimmune MRL/Mp mice.

Congenic mice of the MRL/Mp strain spontaneously develop an autoimmune connective tissue disease that shares immunologic and histopathologic features with the human disorders systemic lupus erythematosus, rheumatoid arthritis, and systemic vasculitis. The autoimmune disorder in these mice is markedly accelerated by the recessive gene lpr. Older MRL/Mp-lpr/lpr mice develop significant inflammatory ocular disease, including choroiditis, scleritis, and orbital vasculitis. Animals of both the MRL/Mp-+/+ and MRL/Mp-lpr/lpr substrains develop lacrimal gland inflammatory infiltrates. The MRL/Mp mouse provides a potential model for ocular inflammatory disease and for Sjögren's syndrome.

Inhibition of conjunctival transdifferentiation by topical retinoids.

During the healing of a total corneal epithelial defect extending beyond the limbus, conjunctival transdifferentiation can be inhibited by corneal vascularization as evidenced by the lack of morphological transformation of the conjunctival epithelium into a cornea-like epithelium and the persistence of goblet cells on the corneal surface. We speculated that corneal vascularization might play a causative role in inhibiting conjunctival transdifferentiation, and examined the hypothesis that vitamin A or retinoids might be one of the blood-borne factors in modulating this process. To test this hypothesis, we created total corneal epithelial defects extending 3 mm beyond the limbus in rabbits using n-heptanol, and segregated the resultant corneas into nonvascularized and vascularized groups. After re-epithelialization, both groups received topical 0.1% Etretinate (Roche-Hoffmann, Nutley, NJ) or 13-cis retinoic acid in corn oil three times a day for 8 weeks. Controls received corn oil only. The extent of transdifferentiation was analyzed by assaying goblet cell density and distribution using flat-mount preparations and Alcian blue and periodic acid-Schiff stains (Fischer Scientific Co., Fair Lawn, NJ) and by conventional histology. Topical retinoid application inhibited conjunctival transdifferentiation in nonvascularized corneas to the same extent as that caused by corneal vascularization, suggesting that vitamin A is an important blood-borne factor for goblet cell maintenance. Its relative deficiency in the normal avascular cornea may explain why conjunctival transdifferentiation occurs.

Endothelial metaplasia in the iridocorneal endothelial syndrome.

PURPOSE: To test the hypothesis that the aberrant, cytokeratin-expressing cells that replace endothelium in the iridocorneal endothelial (ICE) syndrome are of endothelial origin. METHODS: Corneas from four patients with Chandler's syndrome and three with essential iris atrophy were examined by two-color immunofluorescence for simultaneous expression of cytokeratins and two markers of endothelial lineage: vimentin and the antigen recognized by the antiendothelial monoclonal antibody 2B4.14.1. RESULTS: In six corneas, unequivocal endothelial staining for cytokeratins was present; in each of these, cells coexpressing cytokeratins and the two endothelial markers were clearly identifiable. In the remaining cornea, weak cytokeratin staining that colocalized with vimentin was present. CONCLUSIONS: These results lend strong support to the hypothesis that the "epithelial-like" endothelial cells in ICE syndrome are cells of endothelial lineage rather than heterotopia of epithelial cells; these cells probably arise via a metaplastic transformation of preexisting endothelium.

Immunohistochemical pathology of the corneal endothelium in iridocorneal endothelial syndrome.

PURPOSE: To evaluate immunohistochemical staining of the endothelia of corneas from patients with clinical diagnoses of iridocorneal endothelial (ICE) syndrome. METHODS: Corneas diagnosed with ICE syndrome and removed during corneal transplantation were freshly frozen, sectioned, and stained with monoclonal antibodies to keratin subgroups, vimentin, desmin, and a series of other antibodies against intermediate filaments. Transmission electron microscopy was performed on segments of these corneas fixed in glutaraldehyde. RESULTS: There was almost universal staining of the endothelial layer with A1 and A3 keratin monoclonal antibodies and vimentin. Transmission electron microscopy of the corneas also confirmed features consistent with keratin. CONCLUSIONS: The "endothelial" cell layer in the iridocorneal endothelial syndrome has electron microscopic and immunohistochemical characteristics of epithelial-like cells, but it also cross-reacts with vimentin, suggesting that these cells retain or derive some endothelial staining characteristics as well. This "epithelialization" of the endothelial layer may explain the progressive cellular proliferation across angle and iris similar to that seen in iatrogenic epithelial downgrowth and posterior polymorphous endothelial dystrophy.

Goblet cell density and vascularization during conjunctival transdifferentiation.

After debridement of the entire corneal epithelium with n-heptanol, two groups of rabbit corneas were segregated according to the extent of corneal neovascularization. Using a new topographic goblet-cell counting method and routine histology, the authors have reexamined the process of conjunctival transdifferentiation and compared the changes of goblet-cell density and morphology between nonvascularized and vascularized groups for a follow-up period of 167 days. Analysis of the total goblet-cell density disclosed that no goblet cells appeared on the corneal surface during the entire period of reepithelialization. After that, two phases were identified with respect to goblet-cell density: phase I (day 0-17) and phase II (after day 17). In phase I, both groups had a similar surge of goblet cells, with the peak occurring between days 7 and 11, suggesting little correlation with vascularization. Morphologic studies indicated the presence of a prominent centripetal cellular migration. In phase II, the nonvascularized group showed a rapid decline in goblet-cell density, and as a result the morphologic transdifferentiation into a cornea-like epithelium was completed on day 43. The changes of goblet cells to a smaller size and the presence of a more acidic mucin in the centrifugal receding zone, suggested that transdifferentiation on nonvascularized corneas is a process involving changes of cellular differentiation. In contrast, the vascularized group maintained a high plateau of goblet-cell density and an epithelium with conjunctival characteristics until day 167. This result disclosed that retardation of conjunctival transdifferentiation by corneal vascularization was in phase II. The possible role of vascularization in the modulation of conjunctival transdifferentiation is discussed.