Published reports indicate that HIV...

Published reports indicate that HIV is retrieveed from BAL fluid of patients with AIDS who have LIP still not with other AIDS-related pulmonary disease. Our experience has been different. Ten BAL specimens from nine patients with AIDS were cultur directly in peripheral children mononuclear cells, and all ten agricultures were positive for HIV as indicated by way of examination of the culture supernatant from reverse transcriptase assay and enzyme immunoassay for HIV antigen. Five of the specimens were also positive for Pneumocystis carinii, and other pulmonary diagnoses included histoplasmosis, lymphoma, Kaposi's sarcoma, and aspiration pneumonia. Five additional BAL specimens were cultur after freezing at -70[degrees]C yet only two were culture-positive for HIV (p = 0022; FET) This subject of attention indicates that HIV can be win backed from the BAL fluid in principally patients with AIDS, unrelated to the sign of pulmonary disease. In contrast to agricultures HIV antigen was detected in the BAL fluid of solitary one patient, and that patient had LIP with noncaseating granulomas. Therefore, HIV refinement is not useful in the diagnosis of LIP, on the other hand HIV antigen detection should be studied further. All BAL fluids should be considered potentially infectious.

(Chest 1989; 96:64-67)

Isolation of HIV from BAL fluid from patients with AIDS who have LIP has been reported, further not from patients with other AIDS-related pulmonary disease.[1,2] onward this basis, it has been allude toed that the presence of HIV in BAL fluid may be useful in the diagnosis of LIP. These accrues are surprising, since HIV is widespread in carcass fluids of infected patients in the couple T4 lymphocytes and macrophages. united would expect to find HIV in BAL fluid from greatest in quantity patients with AIDS, unrelated to the exemplar of pulmonary disease, because BAL fluid contains the two T4 lymphocytes and macrophages.[3] To investigate this further, a series of BAL fluids from adult patients with AIDS were cultur for HIV and criterioned for HIV antigen.

Materials and Methods

Bronchoalveolar lavage is done routinely in all adult patients with AIDS with persistent respiratory symptoms at the University of Cincinnati. After the report of the usefulness of HIV cultivation in the diagnosis of LIP, BAL fluid was routinely submitted for HIV tillage Ten consecutive bronchoalveolar lavages which were done early enough in the day to allow immediate processing of HIV refinements without overnight storage were studied. An additional five frozen (-70[degrees]C) BAL specimens were cultur for HIV, including common of the fresh specimens which was cultur the pair before and after freezing.

The measure for handling BAL specimens from immunocompromised patients has been described in detail.[4] Bronchoalveolar lavage was performed in the usual manner, from using a fiberoptic bronchoscope (Olympus BF10 or P10) After the patients received topical anesthesia with 2 percent lidocaine, the bronchoscope was introduced into the trachea either transnasally or from one side an endotracheal tube. The bronchoscope was advanced and wedged into a distal airway in the area involved from the infiltrate, as seen onward the chest roentgenogram, or into a subsegment of the right middle lobe or lingula if no infiltrate was noted. After wedging, 60-ml aliquots of sterile nonbacteriostatic saline solution (09 percent NaCl) were instilled by the and of the suction channel of the bronchoscope and subsequently aspirated through hand with a 60-ml, non-Luer-Loking syringe (Becton-Dickinson). The total amount of saline solution instilled between the walls of the bronchoscope was 240 ml The aspirated fluid was mereed and an aliquot used for agriculture and antigen detection.

For HIV civilization human peripheral blood mononuclear confined apartments were separated with a commercial Ficoll-Hypaque preparation (lymphocyte separation medium, Organon-Teknika, Inc) from the buffy coats of random donors from a local life-blood bank. The separated solitary abode; squalids were placed in 75-sq cm flasks at a concentration of 15 x [10sup6] confined apartments per milliliter in 30 ml of RPMI-1640 medium containing 20 percent fetal calf sera (heat-inactivated), penicillin (50 units/ml), streptomycin (50[mu]g/ml) amphotericin B (125[mu]g/ml) and phytohemagglutinin ("medium A") (5[mu]g/ml) The flasks were incubated for three days at 36[degrees]C in 5 percent [COsub2] At this time, 25 ml of the medium was remov from undivided of the flasks, and 5 to 10 ml of BAL fluid was added. Five milliliters of RPMI-1640 medium containing 20 percent inactivated fetal calf sera, 1 percent interleukin-2 (Advanced Biotechnologies, Inc), hexadimethrine bromide (Polybrene) (10[mu]g/ml) anti-human interferon (Interferon Sciences) (40 units/ml), and antibiotics as mentioned earlier ("medium B") was also added, and the agriculture was incubated at 36[degrees]C in 5 percent [COsub2] for undivided hour. Twenty-five milliliters of the same medium was then added and the flask recured to the incubator. After three to four days, 25 ml of the medium was replaced with new medium B; and after seven days, 10 ml of the refinement fluid was removed, the small rooms pelletized (250 x g; 15 minutes) and the supernatant fluid assayed for HIV by means of reverse transcriptase and an HIV antigen assay.[5] The turn topsy-turvy transcriptase assay was considered positive if more than 10000 cpm of activity were fix per milliliter of the civilization fluid. The HIV antigen in the agriculture as well as in the patient's descendants and BAL fluid, was measured with a commercially available enzyme immunoassay (Abbott Laboratories, Inc.) which descrys predominantly p24 antigen. The BAL fluid that was positive for HIV antigen was neutralized with HIV antisera to confirm the specificity. Greater than 50 percent blocking was required for confirmation. Quantitation of antigen was done with standards provided at the Virology Reference Laboratory of the NIAID AIDS Clinical subject of attention Groups. Sampling of cultures was continued until sum of two units consecutive reverse transcriptase assays were positive or for six weeks if the improvement remained negative. During the period of subject of attention none of the uninfected donor enclosed spaces became contaminated with HIV, and solely six (26 percent) of 23 non-BAL specimens were positive.