The safety and immunogenicity of a recombinant outer surface protein A (OspA) Lyme vaccine in patients previously diagnosed with Lyme disease was assessed in a dose-ranging, prospective study. Thirty healthy volunteers were consecutively assigned to receive three doses of 3, 10, or 30 micrograms of OspA vaccine at 0, 1, and 2 months. Subjects were seen 3 days after each vaccine dose and 1 month after completion of the three-dose schedule. Local side effects included soreness, induration, swelling, and redness. Transient systemic side effects occurred in 21 subjects, the majority of which (81%) were characterized as mild. Solicited symptoms included migratory mild arthralgias that lasted 24 h in 3 subjects. Side effects were not more evident after the second or third dose. Of the patients, 93% developed high-titer OspA antibodies. Thus, an OspA vaccine may be safe and immunogenic in patients with a history of Lyme disease.

Cytokineplasts (CKPs) are membrane-bounded, anucleate, granule-poor cytoplasmic fragments, induced from PMNs by brief heat (45 degrees C, 9 min), which retain motile function including chemotaxis and phagocytosis. CKPs can respond to repeated chemotactic stimuli even after having been held overnight at room temperature, and hence "outlive" control PMNs. We now report that adherent CKPs lack significant oxidase activity, as measured by reduction of nitroblue tetrazolium (NBT) dye, (1) 5 min after heat, when they are often still attached to their parent PMNs (which generally do not reduce NBT either); (2) later on, when they are free; and (3) when cells have been pretreated on endotoxin-coated substrata or with phorbol myristate acetate (PMA); both pretreatments cause the large majority of adherent control PMNs to reduce NBT. Moreover, cells harvested from glass just after heat lack the normal increase in oxygen consumption seen on stimulation with PMA or with heat-killed staphylococci. PMA-stimulated respiratory burst activity was not restored to heated cells by exogenous NADPH. Thus, heat applied to normal PMNs can dissociate motile function from oxidase activity; in this respect CKPs resemble PMNs in chronic granulomatous disease. The apparent increased functional stability of CKPs may indicate that normal PMNs are not immune to their own oxidative killing mechanism.

An immunogenic region of the Borrelia burgdorferi flagellin encompassing amino acids 197-273 and designated 41-G was evaluated as an antigen in an enzyme immunoassay (EIA) for Lyme disease on a routine basis in a reference laboratory. Sera that tested positive for Lyme disease by EIA using 41-G or the whole-cell Borrelia burgdorferi lysate as the antigen were also evaluated by immunoblot for reactivity with Borrelia burgdorferi, and the patient's clinical history was determined retrospectively by a questionnaire distributed to the referring physician. The sensitivity of the 41-G based EIA for the serologic diagnosis of Lyme disease, when compared with that of the Borrelia burgdorferi lysate EIA, was 70% (35 of 50). These data demonstrate that 41-G has utility as an antigen in EIA, although the sensitivity is at present less than that of the assay employing the Borrelia burgdorferi whole-cell lysate.

Physiologic concentrations of urate in solution had a clear and dosagedependent suppressive effect on the concomitant uptake of radioiodinated human serum albumin (13'EHSA) during the ingestion of particles (bacteria) by human polymorphonuclear leukocytes (PMN). The effect was rapid, reversible on washing the cells, and demonstrable over a wide range of incubation times and ratios of particles to PMN. In this method, the incubated cells are washed, and most surface radioactivity is thereby removed, the amount of label trapped during particle ingestion then usually provides a sensitive measure of phagocytosis. However, no evidence for an effect of urate on phagocytosis was found by use of several other methods for the assessment of particle u p take, including 1) direct microscopic counts of cell-associated bacteria, 2) studies in which the bacteria were radiolabeled (instead of the media), and 3) the quantitative recovery of surviving bacteria from the media and from disrupted cells. An alternative explanation for the results with '"I-HSA is that urate interferes reversibly with the binding of albumin to the cell membrane, including those portions that become internalized during ~~ From the Section of Rheumatology, Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut.Supported in part by grants from the USPHS (AM-10493, AM-19742, AM-5639, AM-07107), the Arthritis Foundation, and the Kroc Foundation of Santa Ynez, California. Part of this work has appeared in abstract (I).Stephen E. Malawista phagocytosis, thereby creating the false impression of less phagocytosis, when there is really only less trapped label (i.e., albumin). In fact, "on-phagocytizing leukocytes-which do not transport albumin across the cell membraneincubated in serum-bder with added urate and with '"I-HSA and then drained (not washed) had much less cell-associated radioactivity than controls without added urate. Washing removed the difference (and most of the label from both groups). Crystals of urate and of silica are known to kill the cells that ingest them by disrupting the membranes of phagolysosomes, but only after protective protein has been digested. In the presence of dissolved urate, this inflammatory mechanism might be augmented by the relative lack of protective, membrane-associated protein. Indeed, as measured by the release of lactic dehydrogenase, injury to cells given silica crystals was augmented considerably in the presence of dissolved urate.The persistent defect that characterizes gout is the supersaturation of body fluids with sodium urate and the propensity of that urate to form crystals, especially in joints and skin. Superimposed upon the persistent defect are periodic inflammatory paroxysms: acute gouty arthritis. A great deal of work has dealt with how urate crystals, through their interaction with phagocytic cells, contribute to the propagation and amplification of gouty inflammation (for review, see reference 1); one aspect of that interaction-the effect of crystals on membrane i...