Association of HLA types A1-B8-DR3 and B27 with rapid and slow progression of HIV disease

We examined how HLA types A1-B8-DR3 and B27 were related to progression of clinical disease and rate of loss of CD4 lymphocytes in the Edinburgh City Hospital cohort of HIV-positive patients, mainly injection drug users. Patients (n = 692) were prospectively followed from 1985 through March 1994. Accurately estimated seroconversion times were determined retrospectively for a subgroup of 313 (45%). Of 262 patients (39%) who were fully or partially HLA typed, 155 (50%) had known seroconversions. Of 34 patients typed positive for A1-B8-DR3, 29 progressed to CDC stage IV, 22 to AIDS and 20 died. Twelve patients were typed positive for B27; six of these progressed to CDC stage IV, one to AIDS and none died. In a proportional hazards analysis of the 313 patients with known seroconversions, A1-B8-DR3 was significantly associated with covariate-adjusted relative risks of 3.7 (95% CI 1.9- 7.2), 3.1 (1.6-6.0) and 1.9 (1.1-3.2) for progression from seroconversion to death, AIDS and CDC stage IV, respectively. Events for B27 were too rare to include B27 in analyses to death and AIDS, but B27 was significantly associated with slower progression to CDC stage IV (0.3, CI 0.1-0.9). Random effects growth curve models were used to estimate individual rates of loss of square root CD4 count and loss of CD4 percentage, for 603 and 617 patients, respectively. A1-B8-DR3 was associated with rapid loss of both markers (p = 0.02 and p = 0.01, respectively); B27 was associated with slow loss of both markers (p = 0.04 and p < 0.005).      

Risk of HIV infection in recipients of untested blood from donors now anti-HIV-positive

Recipients of untested blood from donors who at a subsequent donation were positive for HIV antibody by enzyme immunoassay (EIA) were evaluated, whether the result on Western blot (WB) assay was negative (EIA+/WB-) or positive (EIA+/WB+). For 109 EIA+/WB- donors, 78 recipients were tested for HIV antibody, and 3 (4%) were positive. Two of the three anti-HIV-positive recipients had clotting disorders, and the other had been massively transfused; in each of these three cases, subsequent test data exonerated the EIA+/WB- donor. For 101 current EIA+/WB+ donors, 35 recipients were tested for HIV antibody, and 13 (37%) were positive. For donors subsequently found to be EIA+/WB+, the rate of isolation of HIV was the same whether the recipients were anti-HIV-positive or anti-HIV-negative (each, 5/6). While recipients of blood from donors subsequently found to be EIA+/WB+ were at substantial risk for HIV infection, regardless of the donor's subsequent HIV culture result, risk of HIV infection was not demonstrated for recipients of blood from donors later found to be EIA+/WB-.     

Epidemiologic characteristics of blood donors with antibody to human immunodeficiency virus

From March 1985 through July 1986, blood donors who were positive for antibody to human immunodeficiency virus (HIV) were evaluated at three major blood centers in the United States. Of 818,629 donations, 450 (0.05%) were HIV antibody-positive. The seroprevalence decreased from 0.07 to 0.04 percent during the study period, due perhaps to a decline in repeat donors. HIV-seropositive donors tended to be 20 to 29 years old (52%) and male (88%). HIV seroprevalence among white donors (2/10,000 donations) was less than that among Hispanic (9/10,000; p less than 0.0001) and black donors (31/10,000; p less than 0.0001). Of 152 seropositive men interviewed, 77 percent reported sexual contact with men; of this latter group, 53 percent were bisexual. Fifteen (44%) of 34 seropositive women had apparently acquired infection from heterosexual contact, and an equal number denied having any known risk factors for HIV infection. Educational efforts must address women and bisexual men who do not perceive themselves to be at risk for HIV infection and should be specifically designed for the mores of different racial and ethnic groups.     

Photodynamic therapy of viral contaminants with potential for blood banking applications

A photodynamic method has been evaluated as a means of eradicating viral contaminants with the potential for rendering blood safe for transfusion. Herpes simplex virus type 1 (HSV-1) was tested under flowing conditions in culture media or in blood supplemented with the virus. Hematoporphyrin derivative was used as the sensitizer and was photoactivated with visible light at 630 nm and 5 J/cm2. HSV-1 in suspension both in culture medium as well as in blood was shown to be killed. The human immunodeficiency virus was also found to be photoinactivated in flowing cell culture medium and, thus, potentially may be inactivated in blood. These findings extend our previous studies which demonstrated that enveloped viruses can be photoinactivated with hematoporphyrin derivative in a static fluid system. Analysis of blood cell number, red cell lysis, plasma proteins, and other standard hematological tests showed no significant change. The possibility that transfusion-associated acquired immunodeficiency syndrome (AIDS) may result from a blood unit infected with human immunodeficiency virus that tested negative makes it imperative that a safe and effective means of viral killing be developed. The system reported here offers promise as an effective approach to this problem.     

T Helper Subset Differentiation in the Absence of Invariant Chain

The outcome of murine infection with Leishmania major is regulated by major histocompatibility complex class II–restricted T helper cells. Invariant chain-deficient (Ii −/−) mice have impaired ability to present major histocompatibility complex class II–restricted antigens, and reduced numbers of CD4⁺ T cells. Despite these deficits, C57BL/6 Ii −/− mice controlled L. major infection comparably to wild-type mice. As assessed by mRNA analysis and in vitro antigen restimulation for IFN-γ, Ii −/− mice had normal induction of Th1 subset differentiation even though antigen-dependent proliferation of their lymph node cells was substantially compromised. In addition, BALB/c Ii −/− mice exhibited a progressive course of infection and Th2 effector cell development that were comparable to that seen in wild-type BALB/c mice. We wished to determine whether this unexpected efficiency of T helper subset induction despite inefficient T cell stimulation could be modeled in vitro. In the presence of rIL-12 or rIL-4 naive parasite-specific transgenic T cells could mature into IFN-γ–or IL-4–secreting T helper cells, respectively, even when antigen presentation was suboptimal or antigen dose was submitogenic. These experiments demonstrate that activation of T helper cells to a threshold required for IL-2 production or proliferation is not required to achieve induction of disease-regulating T helper cell effector functions, and that pathogen-associated secondary activation signals may facilitate the full differentiation of T helper subsets during limiting presentation of antigenic peptides.Experimental infection of inbred strains of mice with Leishmania major remains an exceptional model for analysis of CD4⁺ subset differentiation in vivo (1). Control of disease is dependent on class II–restricted Th type 1 (Th1) cells and their production of IFN-γ which is required to activate macrophages to restrain intracellular replication of the organism. Studies in T cell– (2–4) and IFN-γ–deficient (5) mice have confirmed the critical requirements for these elements in host immunity. MHC class II–deficient mice from a genetically resistant background are completely susceptible to infection (6, 7), while MHC class I–deficient mice from a genetically resistant background retain resistance to infection (8). In contrast to most strains of mice, BALB animals are unable to contain L. major due to the development of an aberrant Th type 2 (Th2) response during infection. The absence of class I does not impact Th2 development or susceptibility in BALB/c mice (9). Leishmania replicates productively only in host macrophages within an endolysosomal-like compartment that contains MHC class II molecules, some of which are devoid of invariant chain (10, 11). Infection of macrophages in vitro is associated with diminished MHC class II–dependent presentation of exogenous antigens (12, 13). Although it is unclear whether this is due to degradation or inappropriate trafficking of MHC class II/peptide complexes from the parasitophorous vacuole (14), competent MHC class II molecules reach the cell surface as demonstrated by immunofluorescent, functional, and biochemical studies (12, 13, 15). Since invariant chain is involved in both targeting newly synthesized MHC class II molecules to peptide-generating compartments, and in protecting the peptide cleft during transit from the endoplasmic reticulum (16), we expected significant impairment in host immune responses to L. major using invariant chain–deficient (Ii −/−) mice. Unexpectedly, both Th1 and Th2 responses were maintained in mice on genetically resistant or susceptible backgrounds, respectively, emphasizing the capacity of the immune system to sustain T cell effector development even under conditions of suboptimal stimulation.     

FGF-2 regulates neurogenesis and degeneration in the dentate gyrus after traumatic brain injury in mice

We studied the role of FGF-2 on regulation of neurogenesis and cell loss in the granule cell layer (GCL) of the hippocampal dentate gyrus after experimental traumatic brain injury (TBI). In both FGF-2(-/-) and FGF-2(+/+) mice subjected to controlled cortical impact, the number of dividing cells labeled with BrdU, injected on posttrauma days 6 through 8, increased at 9 days after TBI, and the number of BrdU-positive cells colabeled with neuron-specific nuclear antigen significantly increased at 35 days. However, in injured FGF-2-/- mice, BrdU-positive cells and BrdU-positive neurons (days 9, 35) were fewer compared with FGF-2(+/+) mice. There was also a decrease in the volume of the GCL and the number of GCL neurons after TBI in both FGF-2(-/-) and FGF-2(+/+) mice, but the decrease in both was greater in FGF-2-/- mice at 35 days. Overexpression of FGF-2 by intracerebral injection of herpes simplex virus-1 amplicon vectors encoding this factor increased numbers of dividing cells (day 9) and BrdU-positive neurons (day 35) significantly in C57BL/6 mice. Furthermore, the decrease in GCL volume was also attenuated. These results suggest that FGF-2 upregulates neurogenesis and protects neurons against degeneration in the adult hippocampus after TBI, and that FGF-2 supplementation via gene transfer can reduce GCL degeneration after TBI.     

Thrombolytic Therapy Within One Hour of the Onset of Acute Myocardial Infarction

Between 1986 and 1988, 239 consecutive patients with acute myocardialinfarction received thrombolytic therapy up to 285 minutes afteronset of symptoms; in 39 (17 with anterior infarction, 21 inferior,one lateral infarction) recombinant tissue plasminogen activatoror anistreplase was administered a mean of 51 minutes (range20–60 minutes) after the onset of symptoms. The patencyrates (26 of 30; 87 per cent), rapid ST segment resolution (36of 38; 95 per cent) and QRS score of 3 (28 of 38; 74 per cent)were statistically significantly higher for those seen and treatedin the first hour in comparison with those seen and treatedlater. For those treated within the first hour mean peak creatine kinasewas 1264 U/I for those with TIMI grade 2 or 3 (partial or completeperfusion) compared with 3005 U/I for those with TIMI grade0 or 1 (no perfusion or penetration without perfusion) (p=0.02):mean peak creatine kinase-MB for those with TIMI grade 2 or3 perfusion was 115 U/I compared with 312 U/I for those withTIMI grade 0 or 1 (p=0.01). Four of the 39 patients developedventricular fibrillation following thrombolytic therapy, threewithin 24 hours of infarction and one following reinfarctionon day 15. There were no significant bleeding complications.One patient died. Thrombolytic therapy within 1 hour of the onset of symptomsled to a very high angiographic patency rate, rapid ST segmentresolution with preservation of left ventricular function. Thistherapy is without significant complications.     

Kisspeptin and GPR54 immunoreactivity in a cohort of 518 patients defines favourable prognosis and clear cell subtype in ovarian carcinoma

Background

Kisspeptins and their G-protein coupled receptor, GPR54 are required for GnRH release and have been associated with anti-metastatic tumour cell behaviour in model systems. The latter might suggest that their overexpression would be associated with a better prognosis in cancer. However, kisspeptin/GPR54 interactions (autocrine, paracrine, and/or endocrine) could also impact tumour behaviour in a negative manner. Here, for the first time, we associate the immunoreactivity of the kisspeptin/GPR54 ligand-receptor pair with favourable prognosis in a large cohort of ovarian carcinomas.

Methods

Immunohistochemical analysis for kisspeptin and GPR54 was performed on a tissue microarray (TMA) consisting of 518 early stage ovarian carcinomas, all with linked clinical outcome data. The TMA was scored using a staining intensity scale of 0 (negative), +1 (mild-moderate), and +2 (strong). Strong staining cases were considered either kisspeptin or GPR54 positive and designated as 1, while all other cases were considered negative and designated 0. All statistical analysis was conducted using two-sided tests and a p value equal to or less than 0.05 was considered significant.

Results

Kisspeptin and GPR54 immunoreactive cases show a favourable prognosis in univariable disease specific survival (p = 0.0023, p = 0.0092), as well as in overall survival (p = 0.0006, p = 0.0002). Furthermore, kisspeptin is an independent marker for favourable prognosis as determined by multivariable disease specific (p = 0.0046) and overall survival analysis (p = 0.0170), while GPR54 is an independent marker for overall survival only (p = 0.0303). Both kisspeptin positive and GPR54 positive cases are strongly associated with the ovarian carcinoma clear cell subtype (p < 0.0001, p < 0.0001), and GPR54 is significantly associated with favourable prognosis in overall survival within the clear cell subtype (p = 0.0102).

Conclusion

Kisspeptin and GPR54 immunoreactivity are significantly associated with favourable prognosis in both disease specific and overall survival, as well as being significantly associated with the clear cell ovarian carcinoma subtype, thereby creating the first independent prognostic biomarkers specific for ovarian clear cell carcinomas.     

T cell clones specific for hybrid I-A molecules. Discrimination with monoclonal anti-I-A (k) antibodies

Alloreactive and soluble antigen-reactive, I-A-restricted T cell clones were examined for their ability to recognize hybrid I-A antigens. Several clones that recognized hybrid I-A(b)/I-A(k) molecules on (C57BL/6 x A/J)F(1) [(B6A)F(1)] spleen cells were studied. We were able to distinguish clones that recognized hybrid I-A molecules of the A(b)(a)A(k)(β) type from those that recognized A(k)(a)A(b)(β) molecules. We reached this conclusion by considering data from three independent types of experiments. (a) Monoclonal antibodies were used to inhibit T cell stimulation. Antibodies 10.2.16 and H116.32 distinguished two mutually exclusive “families” of T cell clones. One group of clones was inhibited by 10-2.16 and not H116.32, the other group exhibited reciprocal inhibition. (b) T cell proliferation was assayed using antigen-presenting cells from B6.C-H-2(bml2) (bml2) and [bml2 × B10.A(4R)]F(1) mice. Because the bml2 strain has a mutation that results in an altered A(b)(β) polypeptide chain (A(bm12)(β)), we reasoned that clones that could recognize the [bm12 × B 10.A(4R)]F(1) cells were recognizing A(b)(a)A(k)(β) molecules. Alternatively, clones not recognizing [bml2 × B10.A(4R)]F(1) cells had specificity for A(k)(a)A(b)(β) molecules. (c) I-A molecules immunoprecipitated from radiolabeled (B6A)F(1) splenocyte extracts were analyzed by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These experiments confirmed an earlier report that antibody 10.2.16 recognized determinants on the A(k)(β) chain (12). Antibody H116.32 immunoprecipitated products consistent with recognition of A(k)(a) determinants. Taken together, these three types of results offer conclusive evidence that T cell clones recognizing “hybrid” I-A molecules use either A(b(k)A(k)(β) or A(k)(a)A(b)(β) molecules as recognition or restriction sites. Clones whose proliferation was supported by [bm 12 x B10.A(4R)]F(1) cells and blocked by anti-I-A(k) antibody 10-2.16 recognized A(b)(a)A(k)(β) B molecules. Clones that were blocked by antibody H116.32 and did not recognize [bml2 X B10.A(4R)]F(1) cells use a recognition site(s) on A(b)(a)A(k)(β) molecules. Thus, we can demonstrate both functionally and biochemically that hybrid F(1) I-A molecules of the structure A(k)(a)A(b)(β) and A(b)(a)A(k)(β) both exist on (B6A)F(1) splenocytes and that both configurations are used in immune recognition phenomena.