Standardization of flow cytometric detection of antigen expression.

Since response to antigen-based immunotherapy relies upon the level of tumor antigen expression we developed an antigen quantification assay using ABC values. Antigen quantification as a clinical assay requires methods for quality control and for interlaboratory and inter-cytometer platform standardization. A single lot of Cytotrol™ Lyophilized Control Cells (Beckman Coulter) used for all studies. The variability in antigen quantification across 4 different instrument platforms in 2 separate laboratories was evaluated. The effect of the antibody clone utilized, importance of custom 1:1 molar ratio (fluorophore to protein, F/P) verses off-the-shelf antibodies, and QuantiBrite PE calibration verses linearity calibration combined with a single point scale transformation with CD4 as reference were determined. Use of single lot control cells allowed validation of reproducibility between flow cytometer platforms and laboratories and allowed assessment of different antibody lots, cocktail preparation, and different antibody clones. Off the shelf antibody preparations provide reproducible estimates of antigen density, however custom 1:1 unimolar antibody preparations should be utilized for definitive measurement of antigen expression.Geometric Mean fluorescent Intensity (GeoMFI) was not comparable across instruments and inter-laboratory. The use of CD4 as the reference marker can minimize variability in ABC values. Comparable antigen quantification is vital in managing patients receiving antigen-based immunotherapy. If this assay is to be utilized in a clinical setting, quality control methods have to be instituted to assure reproducibility and allow validation across laboratories. We have demonstrated that use of a lyophilized cell control is highly valuable in achieveing these goals.

A novel approach for characterization of KSHV-associated multicentric Castleman disease from effusions.

A biopsy of lymphoid tissue is currently required to diagnose Kaposi sarcoma-associated herpesvirus (KSHV)-associated multicentric Castleman disease (KSHV-MCD). Patients showing clinical manifestations of KSHV-MCD but no pathological changes of KSHV-MCD are diagnosed as KSHV inflammatory cytokine syndrome. However, a lymph node biopsy is not always feasible to make the distinction. A pathognomonic feature of lymph nodes in KSHV-MCD is the expansion of KSHV-infected, lambda-restricted but polyclonal plasmablasts. To investigate whether these cells also reside in extra-nodal sites, effusion from 11 patients with KSHV-MCD and 19 with KSHV inflammatory cytokine syndrome was analysed by multiparametric flow cytometry. A distinct, lambda-restricted plasmablastic population (LRP) with highly consistent immunophenotype was detected in effusions in 8/11 patients with KSHV-MCD. The same population was also observed in 7/19 patients with KSHV inflammatory cytokine syndrome. The detection of LRP stratified KSHV inflammatory cytokine syndrome into two clinically distinct subgroups; those with detectable LRP closely resembled KSHV-MCD, showing similar KSHV viral load, comparable severity of thrombocytopenia and hypoalbuminaemia, and similar incidences of hepatosplenomegaly. Collectively, the detection of LRP by flow cytometry can serve as a valuable tool in diagnosing KSHV-MCD. KSHV inflammatory cytokine syndrome with LRP in effusions may represent a liquid-form of KSHV-MCD.

Does impaired executive functioning differentially impact verbal memory measures in older adults with suspected dementia?

The purpose of this study was to examine whether executive dysfunction differentially impacts list-learning and story recall tasks in a sample of older adults referred for suspected cognitive impairment. Older adults (N = 61) with mild cognitive impairment (MCI) or probable mild dementia, and those who did not meet criteria for diagnosis of dementia, were assessed using measures of executive function and verbal memory. Two groups were established based on performance on measures of executive function: (a) the No Executive Dysfunction group (NoED; n = 33) consisted of persons without impairment on any obtained measures of executive function; and (b) the Executive Dysfunction group (ED; n = 28) contained persons with impairment on at least one of the measures of executive function. The two groups were compared on performance on two measures of verbal memory, the California Verbal Learning Test-II (CVLT-II) and the Logical Memory (LM) subtest from the Wechsler Memory Scale-Revised (WMS-R). The NoED group performed significantly better than the ED group on the total learning and short delay free recall trials of the CVLT-II. However, there were no significant differences between the groups on the other indices of the CVLT-II (i.e., long delay free recall, recognition, recall repetitions, recall intrusions, or recognition false-positives) or on the immediate and delayed recall trials of the LM measure. These results support previous research demonstrating the impact of executive dysfunction on the acquisition of and short-delay retrieval of verbal information in older adults with suspected cognitive impairment.

Concurrent validity of abbreviated WAIS-III index scores in geriatric outpatients with suspected dementia.

Assessments of older adults with suspected dementia can be time limited and clinicians might consider using abbreviated versions of measures. The present study examined the concurrent validity of abbreviated WAIS-III index scores in a sample of geriatric patients referred for assessment of suspected dementia (N=43; mean age=63.8 years). All 2-subtest estimates of the Verbal Comprehension, Perceptual Organization, and Working Memory index scores accurately estimated more than 80% of cases within +/-2 standard errors of measurement (S.E.M.), and in most cases, more than 90% of cases were accurate at this level. While none of the 1-subtest estimates of these index scores were as accurate, both of the 1-subtest estimates of the Processing Speed index had high clinical accuracy. Abbreviated versions of the four index scores can be substituted in situations with this clinical population where testing time is limited or a patient fatigues easily.

Concurrent validity of WAIS-III short forms in a geriatric sample with suspected dementia: verbal, performance and full scale IQ scores.

Evaluation of intellectual abilities using the WAIS-III is a common component of neuropsychological assessments. However, clinicians might be interested in administering reliable and valid short forms due to practical and clinical reasons. The present study examined the concurrent validity of eight short forms of the WAIS-III with full form IQ scores in a sample (n=43) of geriatric outpatients referred for assessment of suspected dementia. There were no significant differences between the short and full form VIQ scores at P<.01, while half of the short form PIQ and FSIQ scores were significantly different from their respective full form scores at P<.01. Correlations between short and full form IQ scores ranged from .89 to .99. Seven-subtest short forms were able to accurately estimate over 80% of scores within +/-2 S.E.M.s. This study supports limited use of WAIS-III short forms when conducting evaluations of older adults with suspected dementia.