Human immunodeficiency virus p24 antigen testing in cornea donors

PURPOSE: Testing for the p24 antigen of the human immunodeficiency virus (HIV) may detect early HIV infection in the seronegative window; however, falsely reactive results may occur in cadaver specimens. Although neither the Food and Drug Administration (FDA) nor the Eye Bank Association of America requires p24 testing of cornea donors, many tissue banks using other organs from cornea donors do perform this assay, and the FDA requires that eye banks reject corneal tissue if a reactive p24 assay is reported. We investigated the impact of p24 testing on eye banking and corneal transplantation. METHODS: Two clinical cases and records from the Lions Eye Bank of Delaware Valley (LEBDV) were reviewed retrospectively. RESULTS: Two corneas from the LEBDV were transplanted before the reporting of p24 reactivity by other tissue banks. In one case, because of the young age of the recipient, the surgeon elected to replace the cornea with new tissue hours after the original transplant, and later polymerase chain reaction (PCR) testing was negative. In the other case, there was not enough specimen to perform Western blot or PCR confirmatory testing. The patient was followed with periodic serologic testing for HIV and has remained seronegative. To avoid such problems in the future, the LEBDV initiated testing of all donors with p24 and other nonrequired screening tests. Over a 2-month period, 22 corneas (from 11 donors) were discarded because of these tests: 4 donors had reactive p24 tests, 6 were reactive for antibody to hepatitis B core antigen, and 1 had a reactive syphilis test. CONCLUSIONS: Results from p24 assays by other tissue banks may cause difficult clinical situations when the results are received after transplantation of the tissue, but the use of the p24 assay in the screening of cornea donors may result in excessive waste of donor tissue. Further guidance is needed regarding the management of positive results from this and other nonrequired screening tests.     

Analysis of the serological testing results from Aier Eye Bank北大核心CSCD

Objective: This study was to evaluate the safety of 640 corneal donors by analysing the serological testing results. Methods: We retrospectively analyzed the serological testing results from Changsha Aier Eye Bank and Chengdu Kangqiao Aier Eye Bank from January 2011 to December 2015, hepatitis B virus surface antigen (HBsAg), hepatitis C virus (HCV), treponema pallidum (TP) and human immunodeficiency virus (HIV) were detected by colloidal gold or enzyme-linked immunosorbent assay (ELISA). Results: There were 83 out of 640 serum samples showed positive immuno-reaction assayed markers, the positive rate was 12.97%, including HBsAg(n=60, 9.38%), HCV(n=3, 0.47%), TP(n=11, 1.72%) and HIV(n=2, 0.31%). Moreover, 3 corneal donors were both positive against HBsAg and HCV, 2 donors positive against HCV and TP, 1 donor positive against HBsAg and HIV, 1 donor positive against HBsAg and TP. Conclusions: There is a high proportion of positive results of blood-borne diseases in cornea donors, which is a potential threat to corneal receptors and eye bank workers. Therefore, it is very important to detect serological test strictly for corneal donors. Copyright © 2018 by the Chinese Medical Association.     

Screening cornea donors for antibodies against human immunodeficiency virus. Efficacy of ELISA testing of cadaveric sera and aqueous humor

Coded cadaveric sera from 35 patients with acquired immunodeficiency syndrome (AIDS), from 45 cadavers at high risk of human immunodeficiency virus (HIV) infection, and from 262 cadavers without known signs or risk of AIDS were assessed using three commercially available enzyme-linked immunosorbent assays (ELISA) kits and Western blot analysis. Greater than 94% sensitivity and 99% specificity was achieved with each of the ELISA test kits using cadaveric sera. The Western blot method gave 97.1% sensitivity compared with the autopsy-proven diagnosis of AIDS. Positive results were obtained on sera from AIDS cadavers even if the time of blood draw was delayed 35 hours from death and the time of sera preparation was delayed up to 176 days. False-negative or false-positive ELISA results did not appear to correlate with hemolysis or any parameter of sera preparation. In contrast to the high sensitivity in testing sera, only 16 to 26% of aqueous humor samples from AIDS cadavers were ELISA-positive and 79% were positive by Western blot. These results indicate that three commercially available ELISA test kits are an effective means of screening cadaveric sera for antibodies to HIV, but that aqueous humor cannot be reliably substituted for cadaveric sera to screen potential cornea donors by an ELISA assay.     

Seropositivity of blood samples of 31,355 cornea donors from a tertiary care network of eye banks

International Ophthalmology - To evaluate the seropositivity of human immunodeficiency virus (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV) and syphilis from blood samples of cornea donors...     

HIV antibody screening of corneal donors

Because of the risk of transmitting the acquired immune deficiency syndrome through corneal transplantation, health officials have recommended donor screening. We prospectively studied the seropositivity rate for human immunodeficiency virus infection among ocular tissue donors at our eye bank during 1986. Of 1,517 corneal donors, 5 (0.3%) were repeatedly reactive by enzyme immunoassay. For comparison, 131 (0.06%) of 206,415 blood donors in Houston were similarly seropositive during this same 1-year period. Routine serological testing can be successfully implemented by eye banking personnel to potentially reduce the risk of viral transmission by keratoplasty.     

Incidence of HIV antibody-positive eye/cornea donors in hospital versus medical examiner cases

We retrospectively examined Michigan Eye-Bank and Transplantation Center data on all eye/cornea donors in Michigan and Illinois for whom human immunodeficiency virus (HIV) antibody screening results were available. This was done to compare the HIV-antibody positive serology rate in hospital versus medical examiner donors. This population consisted of 3,783 donor records for the period January 1, 1986 through June 30, 1988--a 30-month period. Of these, 2,628 records were from hospital donors, and 1,155 were medical examiner cases. Of the 2,628 hospital donors, 22 (or 0.83%) tested positive for HIV antibody. Ten of the 1,155 medical examiner donors (or 0.87%) tested positive for HIV antibody. We examined donor demographics of gender, race, age, positive hepatitis B surface antigen, and residence for each group. The difference in the HIV antibody-positive rate between the two groups--hospital and medical examiner donors--is 0.04%, with a slightly higher rate in the medical examiner group. The chi-square was 0.0079, and the p-value was 0.928. We conclude that this difference is neither clinically nor statistically significant.     

Detection of contamination during organ culture of the human cornea

Purpose  

Corneas harvested post-mortem are at risk of contamination, therefore antibiotic additives are used in cold storage and organ culture systems. In the latter, sterility testing of the medium is part of the standard protocol. Intuitively, testing after longer organ culture periods should be more likely to detect contaminations than early testing, but may delay allocation. This study evaluates whether an optimal time for detection of donor cornea contamination can be identified.     

HIV and banked fascia lata.

Concern over the transmission of communicable diseases through donor tissue has recently increased. Nine hundred and fifty-nine pieces of banked homologous irradiated fascia lata have been distributed to ophthalmic plastic surgeons nationwide over the past 3 years since the establishment of the Wills Eye Hospital Fascia Lata Bank. Safeguards taken against the transmission of disease include strict donor selection; negative antibody testing for human immunodeficiency virus (HIV), rapid plasma reagin (RPR), and hepatitis B surface antigen (HbsAg); heat treatment; and radiation sterilization with 4 million rads of cobalt-60 gamma radiation. To date, no cases have been reported of the transmission of HIV through surgical implantation of banked irradiated homologous fascia lata.     

Isolation of the human T-cell leukemia/lymphotropic virus type III from the cornea

Corneoscleral donor tissue from a donor with a positive serum antibody to HTLV-III but without the overt clinical signs of the acquired immune deficiency syndrome (AIDS) was cultured for the presence of the human T-cell leukemia/lymphotropic virus type III (HTLV-III). The virus was isolated from the two corneal specimens in this patient after the tissue had been stored for four days in McCarey-Kaufman medium. The presence of HTLV-III was confirmed by the detection of viral core proteins (approximately 24,000 protein, termed P24 gag), by immunofluorescence of a touch preparation of the corneal epithelium as well as in cells cultured in vitro. The percentage of immunofluorescent cells detected by HTLV-III anti-P24 antibody ranged between 2% and 3%. These findings emphasize the possibility of transmission of this virus via corneal transplantation surgery. Although no documented cases of AIDS have occurred in corneal transplant recipients, serologic screening of donors before the use of the tissue for transplantation is advisable.     

Arbeitsrichtlinien

A cornea/tissue bank must have an organizational structure in which responsibility and authority to issue directives are clearly defined. It must also use a documented quality management system on the basis of good practice procedures which is maintained to the current standards. The personnel of a cornea/tissue bank must be present in sufficient numbers and be suitably qualified. A cornea/tissue bank must be in possession of appropriate facilities which are suitable for the main purpose of preparation of cryopreserved human amniotic membranes from donor placentas. All equipment must be designed and maintained corresponding to the intended purpose. Deviations from the stipulated quality and safety standards must give rise to documented investigations which include decisions on options for correctional and preventive measures. Acquisition of donors and tissue sampling must be strictly controlled and documented. This also applies to entry of donor tissue in the cornea/tissue bank. Cryopreserved human amniotic membranes can only be preserved from donors undergoing caesarean section and who did not present any known infection of the abdominal cavity or any systemic blood borne infection. Contamination of media used for cryopreservation of donor placenta must be ruled out at least once. Measures must be taken to keep the risk of contamination as low as possible. Cryopreserved human amniotic membranes from donor placentas can only be released if defined criteria are fulfilled. Any suspicion of severe undesired reactions and events for the recipient of an amniotic membrane transplant must be registered with the authorities. The activities of a cornea/tissue bank must maintain and adapt to the state-of-the-art with respect to scientific progress.     

Cytomegalovirus replication in cultured human retinal pigment epithelial cells

Retinal pigment epithelium (RPE) was isolated from the globe of a donor positive for human immunodeficiency virus (HIV) who had cytomegalovirus (CMV) retinitis secondary to acquired immunodeficiency syndrome (AIDS). In culture, the cells exhibited normal epithelioid morphology by phase contrast microscopy. After two weeks the cells developed cytomegaly and dense intranuclear and cytoplasmic inclusions and, eventually, died. Transmission electron microscopy (EM) demonstrated intranuclear nonenveloped virus particles 80-120 nm in diameter consistent with a herpes type infection. Immunofluorescence staining demonstrated the presence of CMV antigens. Conditioned medium from the infected cells caused infection in RPE cells isolated from normal donors. Hybridization assay demonstrated the presence of CMV DNA and indicated that the time course of the infection was similar, but not identical to infection in MRC-5 and HEL cells. We conclude that cultured human RPE is a permissive host for CMV.     

Esteraseaktivität eines organotypischen humanen Kornea-Konstrukts (HCC) als In-vitro-Modell für Permeationsuntersuchungen

Organotypic cornea equivalents are used as in vitro models for permeation studies. Many ophthalmic drugs are applied as ester prodrugs to achieve a higher bioavailability. The esterase activity of three corneal human cell lines (epithelial, stromal, endothelial cells) as well as of excised porcine cornea, human donor cornea and human cornea construct (HCC) was investigated and compared. Esterase activity was determined using p-nitrophenyl acetate and hydrocortisone acetate (HCA) as esterase substrates. Hydrocortisone acetate permeation across porcine cornea, human donor cornea and HCC was studied in vitro using Franz-diffusion cells. Corneal epithelial cells showed the highest esterase activity and only small differences to keratocytes and endothelial cells were detectable. The permeation barrier properties of the different corneal tissues were very similar in the case of HCA permeation whereas HCA metabolism rates were in the ranking order of porcine cornea > HCC > human donor cornea.Permeation and metabolism studies indicate that the in vitro permeation model HCC is able to adequately convert hydrocortisone acetate to hydrocortisone.     

Risk of SARS-CoV-2 virus transmission from donor corneal tissue: A review

Since the outbreak of respiratory coronavirus disease (COVID-19) caused by the coronavirus SARS-CoV-2, there is an ongoing discussion about whether the virus could be transmitted through corneal transplantation from donor to recipient. The purpose of this review was to summarize the current knowledge in the scientific community to provide aid in risk evaluation for potential virus transfer by corneal transplants. Literature was searched in PubMed.gov for relevant articles on coronavirus in conjunction with cornea processing, cornea transplantation and eye banking. Further, guidelines of health authorities and eye banking associations were reviewed. Studies have shown that SARS-CoV-2 RNA can be detected in ocular swabs and/or fluid of patients with COVID-19. However, the risk of SARS-CoV-2 virus transmission through these ocular tissues or fluid of patients is judged differently. To date, per literature and official guidelines, no evidence of viable virus in ocular tissue and no cases of transmission of SARS-CoV-2 via tissue preparations have been reported.     

Generation of complement-derived anaphylatoxins in normal human donor corneas

Complement-derived anaphylatoxins (C3a, C4a, and C5a) are potent, stable mediators of acute inflammation. Because human corneas contain functional complement, the authors subjected normal human donor corneas to various forms of immunologic or chemical injury to determine if the complement system could be activated and anaphylatoxins generated. The experimental cornea of each donor pair was injected with lipopolysaccharide (LPS) or immune complexes or injured by application of acid or alkali. The remaining cornea of each donor pair served as a control. After incubation of corneas in tissue culture media for 6 hours and elution in phosphate-buffered saline for 24 hours, C3a, C4a, and C5a were measured in corneal eluates by radioimmunoassay. Compared with control corneas, C3a levels were significantly increased in corneas injected with LPS or immune complexes and in corneas injured with acid or alkali. C4a levels were significantly elevated in corneas injected with immune complexes and in corneas injured with acid or alkali but not in corneas injected with LPS. C5a levels were detectable only in corneas injured with acid or alkali. These results suggest that immunologic reactions in the human cornea may activate the classic or alternative complement pathways and generate anaphylatoxins. Additionally, chemical injuries with acid or alkali generate anaphylatoxins in the cornea. Anaphylatoxins may participate in the acute inflammatory response of the human cornea to chemical or immunologic injury.     

Virologic considerations in donor selection for allogeneic keratoplasty

Over a period of 14 months (from October 1985 to December 1986) 152 potential cornea donors were evaluated for human immunodeficiency virus (HIV) and hepatitis B surface (HBs) antigen. In three clinically normal donors (two deaths due to natural causes, one as a result of an automobile accident), HIV seroconversion was found by ELISA techniques in two cases and a positive Western Blot test in one. The two elderly patients had previously undergone multiple blood transfusions during major abdominal surgery in smaller community hospitals before blood donors were routinely tested for HIV; one 20-year-old patient was probably a homosexual, as in-depth interviews with his family doctor revealed. Hepatitis B serology was positive in four potential cornea donors. The authors conclude that interdisciplinary work-ups of potential cornea donors should include screening for infectious diseases.     

Detection of human immunodeficiency virus, hepatitis B virus, and hepatitis C virus in donor eyes using polymerase chain reaction.

Detection of human immunodeficiency virus (HIV), hepatitis B virus (HBV), and hepatitis C virus (HCV) in donor eyes was performed. DNAs were extracted from the uvea, and they were amplified using the polymerase chain reaction (PCR). Amplified viral DNAs were detected with liquid hybridisation and chemiluminescent assay in which no radioactive materials were used. This method was shown to have a sensitivity limit of fewer than 10 copies of HIV, making it much more sensitive than the current techniques employed in eye banks. The method was applied to 120 donor eyes, including four from donors seropositive for HBV. The HBV gene was detected in one case in which the donor's blood had not been tested for HBV. HIV and HCV genes were not detected in any of the samples. The assay could be an effective screening test for the detection of these viruses in eye bank eyes.     

Preparation of amniotic membrane for ocular surface reconstruction

We describe the preparation and preservation of human amniotic membrane required for transplantation in the management of ocular surface diseases. Informed consent is obtained and the donor is screened to exclude risk of transmissible infections such as human immunodeficiency virus (HIV), hepatitis B virus, hepatitis C virus, and Treponema pallidum infections. Ideally, the media and washing solutions needed for the preparation of amniotic membrane are prepared only a week to 10 days prior to use and not stored in the freezer weeks ahead. The AM obtained under sterile conditions after elective caesarian section is washed free of blood clots and chorion. With the epithelial surface up, amniotic membrane is spread uniformly without folds or tears on individually sterilized 0.22 micron nitrocellulose membranes of the required sizes. The prepared filter membrane with the adherent amniotic membrane is placed in the preservative medium and stored at -80 degrees C. The membranes are released when the repeat serology for HIV after the window period has excluded virus infection in the donor. Depending on consumption they may be used up to 6 months after preparation, though many have recommended storage for an indefinite period. Since the amniotic membrane has only incomplete expression of HLA antigens and amniotic epithelial cells do not express them, it is not rejected after transplantation. The presence of several cytokines in the amniotic membrane promotes epithelialization with reduction of fibrosis during healing.     

Toward electron-beam sterilization of a pre-assembled Boston keratoprosthesis

PurposeTo evaluate the effects of electron-beam (E-beam) irradiation on the human cornea and the potential for E-beam sterilization of Boston keratoprosthesis (BK) devices when pre-assembled with a donor cornea prior to sterilization.MethodsHuman donor corneas and corneas pre-assembled in BK devices were immersed in recombinant human serum albumin (rHSA) media and E-beam irradiated at 25 kGy. Mechanical (tensile strength and modulus, and compression modulus), chemical, optical, structural, and degradation properties of the corneal tissue after irradiation and after 6 months of preservation were evaluated.ResultsThe mechanical evaluation showed that E-beam irradiation enhanced the tensile and compression moduli of human donor corneas, with no impact on their tensile strength. By chemical and mechanical analysis, E-beam irradiation caused a minor degree of crosslinking between collagen fibrils. No ultrastructural changes due to E-beam irradiation were observed. E-beam irradiation slightly increased the stability of the cornea against collagenase-induced degradation and had no impact on glucose diffusion. The optical evaluation showed transparency of the cornea was maintained. E-beam irradiated corneal tissues and BK-cornea pre-assembled devices were stable for 6 months after room-temperature preservation.ConclusionsE-beam irradiation generated no detrimental effects on the corneal tissues or BK-cornea pre-assembled devices and improved native properties of the corneal tissue, enabling prolonged preservation at room temperature. The pre-assembly of BK in a donor cornea, followed by E-beam irradiation, offers the potential for an off-the-shelf, ready to implant keratoprosthesis device.     

Human immunodeficiency virus-positive patient with bilateral corneal endothelial deposits.

BACKGROUND: Ocular disorders associated with the human immunodeficiency virus are numerous as are ocular side effects from medications used to treat all of the manifestations of the virus. This report presents a unique case of bilateral, peripheral, and corneal endothelial deposits that may be a result of either the human immunodeficiency virus or the medication rifabutin. Rifabutin was the only medication prescribed that is known to cause endothelial deposits. Rifabutin is part of a multidrug therapy to prevent or treat Mycobacterium avium complex, a common pulmonary disease of immunocompromised individuals. CASE REPORT: A 69-year-old man with a 20-year history of being human immunodeficiency virus-positive presented with bilateral, asymptomatic, peripheral, and corneal endothelial deposits of unknown etiology. Literature research suggested that the deposits did not appear like cytomegalovirus retinitis-related deposits but rather a variant of rifabutin-associated deposits. CONCLUSIONS: These rifabutin-associated deposits differed from known rifabutin-associated deposits previously reported in the literature. These deposits have increased in pigmentation and density 5 years after the patient discontinued the drug. This case may represent another variation of rifabutin-associated endothelial deposits. Knowledge of human immunodeficiency virus and all the associated ocular findings (owing to both the condition and its treatment) is important, because the length of time patients are living with human immunodeficiency virus is increasing.     

Histology after lamellar keratoplasty and corneal excimer laser ablation.

PURPOSE: To correlate clinical and histological findings after lamellar keratoplasty, phototherapeutic keratectomy, and application of a donor lenticule on a human cornea. METHODS: A cornea was obtained during penetrating keratoplasty. The specimen was fixated, dehydrated and embedded in Epon resin. The tissue was cut in 0.5-microm-thick semi-thin sections, stained with toluidine blue, and studied with light microscopy. RESULTS: The central part of the photoablated cornea, which was covered by the donor lenticule, did not differ from a normal cornea. Peripherally, a hazy ring was found clinically. Histology showed an irregular epithelium. Where it was thickened, the epithelium was hyperplastic and showed an increased number of cell layers. In the hazy region, Bowman's layer was absent, indicating that the donor lenticule did not cover this part of the photokeratectomized cornea. The anterior-most part of the corneal stroma was vacuolized and contained amorphous extracellular material; swollen keratocytes were present in this region. Beneath this layer, collagen lamellae were wavy and interwoven and keratocytes were increased in number, appeared swollen, and some had assumed an atypical shape. Peripheral to the haze, the cornea was clear. Histologically, the epithelium was irregular and hyperplastic, Bowman's layer was absent, and stromal collagen lamellae were abnormally organized, but no vacuolization was found. CONCLUSIONS: The formation of haze after excimer laser photokeratectomy can be minimized if the ablated stroma is covered by a corneal lenticule.