Erik Lucht

Induction of mucosal IgA by a novel jet delivery technique for HIV-1 DNA.

Novel ways of delivering plasmid DNA to elicit humoral IgA, IgG and cell-mediated immune responses in mice were investigated. Intraoral administration of DNA in the cheek, using a jet immunization technique, elicited the highest IgA mucosal responses. Intranasal immunization gave strong mucosal IgA responses and persistent systemic IgG. Immunoglobulin isotype analysis revealed an IgG1 profile for intramuscular tongue and gene gun immunizations and an IgG2a profile following oral jet injection and intranasal application. The route of delivery was of importance for the characteristics and quality of the mucosal immune response following DNA immunization. For DNA vaccine delivery, the intraoral jet injection technique has the advantages of being a simple and rapid way of administering the DNA in solution and of provoking specific mucosal IgA when administered in the mucosal associated lymphoid tissue.

Shedding of Cytomegalovirus and Herpesviruses 6, 7, and 8 in Saliva of Human Immunodeficiency Virus Type 1—Infected Patients and Healthy Controls

We used the polymerase chain reaction to study the presence of DNA from cytomegalovirus (CMV) and human herpesvirus (HHV)-6, HHV-7, and HHV-8 in saliva from 44 human immunodeficiency virus (HIV) type 1-infected patients at different stages of disease and in 15 healthy HIV-seronegative controls. CMV DNA, HHV-6 DNA, and HHV-7 DNA were found in all groups, but HHV-8 DNA was found only in symptomatic HIV-1-infected patients (5 [17%] of 29). One of the patients with HHV-8 DNA in saliva had oral Kaposis sarcoma at the time of sampling, and another later developed the tumor. CMV DNA was found most often in the patients with AIDS. HHV-6 shedding tended to be less frequent among HIV-1-infected patients than among healthy controls. HHV-7 DNA was detected least frequently in the group of patients with AIDS. The presence of viral DNA was not correlated either with antiherpesvirus drug therapy or with oral symptoms, apart from Kaposis sarcoma.

Direct identification by PCR of EBV types and variants in clinical samples

Both Epstein‐Barr virus (EBV) type A and type B, and variants of type A, were identified simultaneously by polymerase chain reaction (PCR) amplification of a DNA region coding for a 13 amino acid repeat in the Epstein‐Barr virus nuclear antigen (EBNA) 6. Whereas this region varies extensively in type A isolates, no variation was seen in type B isolates. When a repetitive region in the LMP1‐coding region was amplified by PCR, it was possible to distinguish individual variants of type B isolates from each other. Forty‐two saliva samples from HIV‐1‐carrying individuals were examined for the presence of type A and type B virus. Both types and multiple variants of each type were found with a much higher frequency than in the saliva samples from healthy individuals. Type A EBV alone was detected in mouthwash samples from 6 infectious mononucleosis (IM) patients. Both type A and B were detected in the peripheral blood B‐lymphocytes (PBL) from 1 healthy individual. The same type A variant was demonstrated both in PBL and in the mouthwash sample from another healthy individual. In this study it was shown that a combination of the EBNA 6‐ and LMP 1‐specific PCRs followed by Southern hybridisation can be used to identify both type A and type B virus, as well as to distinguish between multiple variants of the same strain, in saliva and B‐cells from both healthy and immunosuppressed individuals. J. Med. Virol. 51:355–363, 1997.

DNA-plasmids of HIV-1 induce systemic and mucosal immune responses.

Abstract DNA-based immunization has been shown to induce protective immunity against several microbial pathogens including HIV-1. Several routes of DNA vaccination have been exploited. However, the properties of the immune responses seem to differ with the different routes used for DNA delivery, ultimately affecting the outcome of experimental challenge. We measured the primary immune response following one vaccination. This report presents differences associated with three different DNA delivery routes: intramuscular injection, intranasal application, and gene-gun based immunization. Induction of systemic humoral immune responses was achieved most efficiently by either intranasal or gene-gun mediated immunization, followed by intramuscular injection. Mucosal IgA was reproducibly induced by intranasal instillation of the DNA, and found in lung washings, faeces, and vaginal washings. Cytotoxic T cells were not induced by a single immunization, but were observed after three immunizations using intramuscular injections.

Entamoeba gingivalis in Human Immunodeficiency Virus Type 1-Infected Patients with Periodontal Disease

Necrotic periodontal disease is a progressive painful oral lesion in human immunodeficiency virus type 1 (HIV-1)-infected patients, and the etiology is unknown. Earlier studies of HIV-1-infected patients have shown significant changes in the viral and fungal oral microflora. The aim of this study was to relate the occurrence of protozoa to clinical symptoms and immunosuppression. Oral symptoms were registered in 45 patients at different stages of the HIV-1 infection and in 15 HIV-seronegative healthy controls. Saliva and dental plaque were analyzed for the presence of protozoa. Entamoeba gingivalis was the only protozoa found in the oral cavities of HIV-1-infected patients with periodontal disease. Its presence was not related to the degree of immunodeficiency but to the HIV diagnosis. This study describes for the first time the findings of E. gingivalis in the oral cavities of HIV-1 infected patients.

Epstein-Barr virus (EBV) DNA in saliva and EBV serology of HIV-1-infected persons with and without hairy leukoplakia.

Secretion of Epstein-Barr virus (EBV) in saliva, as well as serum antibody titres against various EBV antigens, were analyzed in respect of (1) 15 HIV-1-infected patients with oral hairy leukoplakia proven to contain EBV by in situ hybridization, (2) 45 HIV-1 infected patients without hairy leukoplakia, (3) 10 HIV-1 infected patients treated with acyclovir or foscarnet and (4) 21 healthy controls. The numbers of CD4+ cells in the peripheral blood were also recorded. The HIV-1 infected patients were at various stages of HIV-1-associated disease. Excretion of EBV DNA in the saliva was determined by means of the polymerase chain reaction (PCR) while the amount of EBV DNA in positive samples was estimated by repeated titrations. The frequency of shedding of EBV DNA increased from 33% in healthy controls to 78% in asymptomatic HIV-1 infected persons, but did not increase significantly with progression of HIV-1-associated disease. The titres of EBV DNA in saliva correlated inversely and significantly with the number of CD4+ cells in the peripheral blood. All patients with hairy leukoplakia shed by EBV DNA in their saliva but the titres were not significantly higher than those of other HIV-1 infected persons. The serum titres of antibodies against EBV nuclear antigen 1 (EBNA-1) correlated positively and significantly with the CD4+ cell count in the peripheral blood. EBNA-1 IgG antibody in the serum was also significantly lower in symptomatic than in asymptomatic HIV-1 infected persons. There were, however, no significant differences in serum antibodies to various EBV antigens between patients with and without hairy leukoplakia.

Opportunistic oral infections in patients infected with HIV-1

Several organs are severely affected in human immunodeficiency virus type 1 (HIV-1) infection. One of these is the oral cavity, in which a variety of lesions of known and unknown causes are observed. The possible causes of some oral lesions and the presence of bacteria, virus and yeasts in saliva and gingival crevices have been investigated in patients infected with HIV-1. Common oral symptoms, which are strongly over-represented in adult patients infected with HIV-1, are Candida infection, necrotic periodontal disease, hairy leukoplakia, dry mouth problems, oral Kaposis sarcoma and oral ulcers. These symptoms are seen mostly in immunocompromised HIV-1-infected patients with other HIV-1-related symptoms. Other less common oral lesions in adult patients infected with HIV-1 are infection with herpes simplex virus, varicella-zoster virus and papilloma virus, malignant lymphoma, salivary gland disease, drug-related ulcers or ulcers infected with microorganisms such as mycobacteria and cytomegalovirus. Changes in the bacterial flora and HIV-1 itself in the oral cavity do not explain the observed lesions. Fungal and viral infections are common in HIV-1-infected patients. Epstein-Barr virus is associated with oral hairy leukoplakia, and cytomegalovirus (CMV) may be correlated to some oral symptoms such as necrotizing gingivitis and dry mouth problems. The shedding of Epstein-Barr virus may indicate early deterioration of the cellular immune response, while CMV appears to be related to late HIV-1-induced symptoms.

Immunoglobulin G abnormalities in HIV-1 infected individuals with lymphoma.

BACKGROUND Polyclonal B-cell activation precedes the occurrence of malignant B-cell clones. Several recent reports suggest a perturbed cytokine regulation in HIV-related lymphomagenesis and Epstein-Barr virus (EBV) involvement in approximately half of the cases with generalized lymphoma. OBJECTIVES We investigated whether altered immunoglobulin properties would be detected by fine analysis of the immunoglobulin G (IgG) subclass patterns against HIV and EBV epitopes. STUDY DESIGN HIV-1 infected patients in early stage, late stage and with lymphoma were analyzed by ELISA for anti HIV and EBV IgG class and subclass antibodies. Avidity and affinity of the antibodies were studied. The lymphoma patients were also studied by PCR for EBV DNA in serum. RESULTS The total IgG reactivity to several HIV antigens was similar in the three patient groups. However, lymphoma patients had a more restricted subclass pattern with significantly lower IgG1 and IgG3 anti gp120 titers compared to other HIV-infected patients but good and persistent total IgG and IgG1 (excluding the gp120 antigen) reactivities in contradiction to their low CD4 counts. IgG4 reactivity was sparse, detectable to significant levels in the symptomatic group only. The observed relative affinity of the HIV-specific IgG and IgG1 of lymphoma patients was similar to that of asymptomatic and symptomatic patients. The subclass reactivity to the EBV peptide was similar in all groups but lymphoma patients with EBV DNA in serum exhibited significantly lower anti EBV peptide titers than those who were EBV DNA negative. CONCLUSION These findings indicate that subclass analysis to defined viral antigens may be a means to detect immune dysregulation in tumor development.