Granulocytes sample from

The ELISA for detection of the DNA adduct was successfully applied to blood samples from two casualties of the Iraq-Iran conflict. These samples were collected 22 and 26 days following the alleged exposure to sulfur mustard (13). Concentrations found in lymphocytes and granulocytes were equivalent to similar levels found in human blood after treatment in vitro with 0.015-0.43 xM sulfur mustard. 

As discussed in Chapter 3, proteins and mABs distribute initially into the plasma volume and then more slowly into the interstitial fluid space. It can be seen from Table 32.11 that the initial distribution volume of interleukin-2 (IL-2) IL-12/ granulocyte colony-stimulating factor (G-CSF)/ and recombinant tissue plasminogen activator (rt-PA) approximates that of plasma volume. In contrast/ the initial distribution volume of FIX is approximately twice that of plasma volume. On the other hand/ the volumes of distribution at steady state (Vd(ss)) for IL-12/ G-CSF/ and rt-PA are considerably smaller than is the Vd(ss) of inuliii/ a marker for extracellular fluid space (ECF). When distribution volume estimates are much less than expected values for ECF/ they could reflect the slow transport of large molecules across membranes and the fact that either assay sensitivity or sampling time has been inadequate to characterize the true elimination phase of the compound. 

The blood from two Iranian casualties that were believed to have been exposed to sulfur mustard in 1988 was analyzed using both the ELISA method for DNA adducts and the GC-MS method for the analysis of the N-terminal valine of hemoglobin (Benschop et al., 1997). Samples were collected 22 and 26 days following the suspected exposure to sulfur mustard. One of the casualties had injuries to the skin that were consistent with an exposure to sulfur mustard, but the second casualty had injuries that were described as only vaguely compatible with sulfur mustard exposure. Both individuals had approximately the same level of hemoglobin valine adduct that was equivalent to the amount observed from a 900 nM in vitro sulfur mustard exposure in whole blood. ELISA DNA adduct levels observed in the granulocytes were also similar for both individuals, 150-160 nM. The individual with the skin injuries consistent with sulfur mustard exposure had observed ELISA DNA adduct levels in the lymphocytes that were only about half that observed in the individual with injuries that were less pronounced, 220 and 430 nM, respectively. 

Leukocyte esterase is released from lysed granulocytes in the urine its presence is suggestive of urinary tract infection. If the processing of the urine sample is delayed, a false-positive leukocyte esterase may be reported if the sample is contaminated. 

Changes in DNA supercoiling were analyzed using nucleoid sedimentation as previously described and are shown in Fig. 1 (2). Human granulocyte-macrophage preciusor cells were prepared from bone marrow samples (obtained with consent from healthy volunteers) by density fractionation and adherence (2). Cells were exposed to two inducers of granulocytic differentiation (retinoic acid and G-CSF) and two inducers of monocytic differentiation (phorbol myristate acetate and vitamin D3). Treatment with differentiation inducers and culture conditions are described elsewhere (2). 

Extreme care should be taken to prevent contamination of the analyser with cell debris. This can arise because of the lysis of granulocytes in samples examined after thawing from oyopreservation or after T ceU depletion. Blockages can be removed by flushing the system with 20-50% (v/v) bleach followed by at least six washes with clean water. If this fails the system can be washed through with 0.1% sterile tiypsin (Sigma Aldrich) fbllowed by further washing with bleach and clean water. 

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