Characterization of immunostimulating complexes (ISCOMS) of HIV-1.

HIV-1, strain HTLV-III, propagated in H9 cells and purified by sucrose gradient centrifugation, was used as native antigen source for the preparation of immunostimulating complexes, HIV-iscoms. The major antigen detected in the iscom was the cell-derived HLA-DR, which readily could be removed from the virus lysate by immunosorbent. In the iscoms the HIV structural proteins MA p17, p55 and TM gp41 were identified; SU gp120 was present in only minute amounts in the virus lysate. The iscom particles appeared well preserved after freeze drying with a round shape, approximately 35 nm in diameter, comprising morphological subunits, assembled with icosahedral symmetry. Immunization experiments in mice reflected the antigen content of the iscoms. High antibody response was induced to HLA-DR in non-depleted iscoms. Major humoral responses were observed to the viral structural proteins MA p17, CA p24, p55, and also to TM gp41. A low or negligible antibody response to SU gp120 was induced by the HIV-iscoms. The negligible response was, however, overcome by the addition of recombinant gp160 to the virus lysate prior to formation of iscoms, resulting in a preparation evoking a clear serum antibody to gp160.

Deletion of the GPG motif in the HIV type 1 V3 loop does not abrogate infection in all cells.

The three amino acids glycine, proline, and glycine (GPG) constitute a conserved motif at the center of the V3 loop of HIV-1 surface glycoprotein 120. It has been indicated that deletion of this GPG motif is lethal for viral infectivity and abrogates the ability of the virus to form syncytia. In the present work, we studied the effects of GPG deletion on viral infectivity, cell tropism, syncytium formation, and initiation of apoptosis by constructing a mutant provirus based on the infectious clone pBRu-2. Successful infection and replication of GPG-deleted virus were detected in MT-2 cells, although the mutant virus showed lower infectivity. Infection could also be observed in the C8166, C91-PL, Molt-3, and THP-1 cell lines, and in PBMC-derived dendritic cells (DCs), but not in CEM-SS, HUT78, H9, Jurkat, and U937 cell lines or in PBMCs. Mutant virus also induced syncytia and apoptosis in the MT-2 cells. An intact GPG motif is probably necessary for unimpaired induction of fusion in some HIV-1-permissive cells. However, once the virus enters the cells, the GPG sequence does not seem to be indispensable for syncytium formation or apoptosis induction in MT-2 cells. Our data also imply that cell surface molecules other than CD4 and CXCR4 may be involved in entry of the GPG-deleted virus.

Spatial visualization of the maturing HIV-1 core and its linkage to the envelope.

The core of late states of maturing human immunodeficiency virus type 1 (HIV-1) has been visualized in three dimensions at approximately 7 nm resolution by electron microscopic tomography. After budding, approximately 25 nm thick precursor material is observed densely assembled inside the viral envelope. Upon proteolysis the core material is transported and condensed in the center of the virion. The core, 100 nm in length, spans the entire diameter of the virion showing a 40-60 nm wide free end and a narrow end approximately 20 nm. A model of the core is derived consisting of two fibers packed into a bilateral, elongated structure. Two ends of the fibers are compacted together, forming one narrow end of the core, while the two other fiber ends are situated more loosely together allowing for flexibility. Structural maturation of the virus could be reflected by the degree of compactness of the core. The narrow end of the core is observed attached to the envelope with a conspicuous core-envelope link (CEL).

Change of occupation and retirement among Swedish farmers and farm workers in relation to those in other occupations. A study of "elimination" from farming during the period 1970-1988.

A number of studies carried out in different countries have shown that farmers have a low morbidity and mortality in comparison to those in other occupations. However, this has been questioned on the basis that some type of selective process may be operating, in that persons having health problems will avoid farming, or are forced to leave farming for other occupations. To determine the occurrence of a so-called 'healthy worker effect', this postal survey of 'elimination' from farming and farming-associated occupations has been carried out. A total of 1283 male farmers and 334 male farm workers born in 1935 and active in Sweden in 1970 were taken as the study group. As controls, a similar number of occupationally active men of the same age and living in the same municipalities were randomly chosen. The results showed that farmers changed occupation or retired early less often than those in other occupations did, whereas more farm workers changed occupation and retired than did other workers of the same age. Among the different reasons given for work change/retirement, low income/poor earning capacity was more common among the farmers and farm workers than among the controls. Illness was less common among farmers but tended to be more common among farm workers as cause of work change. Few farmers changed their occupation because they were offered other work, in comparison to those in other occupations. Allergic disease more often led to an occupation change among farmers, while they less often gave cardiac disease and locomotor problems as a reason for change of occupation, this was probably also true for the farm workers.(ABSTRACT TRUNCATED AT 250 WORDS)

Iron status in athletes involved in intense physical activity.

Iron status was studied in 8 long distance runners. Training programs, diets, and general health were also investigated over a period of at least 2 years. Hemoglobin and serum iron did not differ from normals whereas bone marrow showed either an absence or only traces of iron indicating latent iron deficiency. Radio iron studies of heme and non-heme iron absorption and incorporation showed rather low values. However, they did not differ significantly from normal values. Whole body counting studies during 10 months showed increased elimination of radio iron corresponding to 2 mg iron/day. Due to a high daily iron intake, a balance was present. No signs of bleeding, hemolysis, or increased excretion of hemoglobin via the urine could be found. The low aborption and increased elimination of iron seems to be the explanation for the empty iron stores.

Spatial visualization of progressive states of maturing lentivirus.

Progressive states of maturing lentivirus: maedia visna virus (MVV) and human immunodeficiency virus (HIV), respectively, have been visualized by 2-D electron microscopy and by 3-D electron microscopic tomography. A major fraction of MVV and a low percentage of HIV appear as immature particles 4 to 5 days post virus infection. Upon budding the gag-precursor material is densely packed inside the external envelope. After virus release the major portion of precursors is assembled within an approximately 25 nm thick layer directly attached to the envelope. Structural maturation of the core is different for the two viruses. Pleomorphic cores are observed in mature MVV in contrast to structurally defined cores of HIV. The latter are principally cone-shaped, spanning the entire diameter of the virion with a 40 to 60 nm wide free end and an approximately 20 nm narrow end attached to the envelope with a core-envelope-link.

Inflammatory response and antigen localization following immunization with influenza virus ISCOMs.

The inflammatory response, antigen retention, and antigen localization was studied in mice after immunization with influenza virus glycoproteins presented in two physically defined forms--micelles and ISCOMS (immunostimulating complexes). Two hours after intraperitoneal injection, the proportion of polymorphonuclear leukocytes (PMNs) in peritoneal lavage cells increased from less than 1% to 82% in ISCOM-treated mice and from less than 1% to 41% of the total cell count in micelle-treated mice. For both treatment groups, the proportion of PMNs returned to around zero 24 h postimmunization. Total recovery of radioactive antigen was significantly greater (P less than 0.05) in ISCOM-treated than in micelle-treated mice at one, two, and eight days postinjection. At all times tested, animals given ISCOMs had significantly more radioactive antigen in their spleens than animals given micelles. By electron microscopy ISCOMs were found to attach externally to the plasma membrane or within phagosomes of macrophages in close association with the membranes.

Exposure to microorganisms, febrile and airway-obstructive symptoms, immune status and lung function of Swedish farmers.

A questionnaire was sent to 512 farmers, members of a local farmer's health association. Eighty were interviewed, and their serum precipitins and total serum immunoglobulin E (IgE) values were determined. Forty-five underwent extensive pulmonary function tests. On the basis of the clinical evaluation it was estimated that 19% of the farmers had experienced febrile reactions (fever and/or shivering) following exposure to organic dust, 50% of the 19% having been exposed within the last 2.5 years. Common causes were moldy grain, hay, and woodchips. The pulmonary function, gas exchange, and chest radiographs of those who had previously had febrile reactions were normal. Only 13% showed positive precipitin reactions in Ouchterlony double-diffusion tests. With more sensitive tests, positive precipitins were found in 59%, but they were negatively correlated with febrile episodes. Air samples collected during work with hay and grain on 21 farms contained between 10(7) to 2 X 10(9) microorganisms/m3. The highest values were associated with symptoms of alveolitis. Eleven percent of the farmers reported obstructive chest symptoms following exposure to organic dust. This group showed decreased pulmonary function and elevated serum IgE levels and included a high proportion of smokers and exsmokers. No correlation was found between febrile and obstructive reactions.

Morphology of A/WSN influenza virus-infected chick embryo cells.

A/WSN (H0N1) influenza virus-infected chick embryo cells (CEC) attained a swollen central region 4 hours post infection (p.i.), different from the elongated uninfected cell. From 6 hours p.i. of CEC an increasing number of approximately 100-120 nm, round protrusions were observed by scanning and transmission electron microscopy on the cell membrane, which was smooth on the uninfected cell. At 48 hours p.i. a great number of protrusions were observed on the cell; in addition, there were a number of holes or invaginations in the membranes of the infected cells indicating cell destruction.