Analysis of IgE heavy chain sequences from a small volume of peripheral blood from a mildly atopic individual

2002 ◽  
Vol 109 (1) ◽  
pp. S112-S112
Author(s):  
William JJ Finlay ◽  
Carl S Goodyear ◽  
Jay Slater
Keyword(s):  
Heavy Chain ◽  
Peripheral Blood ◽  
Small Volume ◽  
Atopic Individual

The diversity of rearranged immunoglobulin heavy chain variable region genes in peripheral blood B cells of preterm infants is restricted by short third complementarity-determining regions but not by limited gene segment usage

Blood ◽  
2001 ◽  
Vol 97 (5) ◽  
pp. 1511-1513 ◽  
Author(s):  
Michael Zemlin ◽  
Karl Bauer ◽  
Michael Hummel ◽  
Sabine Pfeiffer ◽  
Simone Devers ◽  
...  
Keyword(s):  
B Cells ◽  
Preterm Infants ◽  
Heavy Chain ◽  
Peripheral Blood ◽  
Gene Segment ◽  
Immunoglobulin Heavy Chain ◽  
Term Infants ◽  
Extremely Preterm ◽  
Extremely Preterm Infants ◽  
Complementarity Determining Regions

The immunoglobulin diversity is restricted in fetal liver B cells. This study examined whether peripheral blood B cells of extremely preterm infants show similar restrictions (overrepresentation of some gene segments, short third complementarity-determining regions [CDR3]). DNA of rearranged immunoglobulin heavy chain genes was amplified by polymerase chain reaction, cloned, and sequenced. A total of 417 sequences were analyzed from 6 preterm infants (25-28 weeks of gestation), 6 term infants, and 6 adults. Gene segments from the entire VHand DH gene locus were rearranged in preterm infants, even though the DH7-27 segment was overrepresented (17% of rearrangements) compared to term infants (7%) and adults (2%). CDR3 was shorter in preterm infants (40 ± 10 nucleotides) than in term infants (44 ± 12) and adults (48 ± 14) (P < .001) due to shorter N regions. Somatic mutations were exclusively found in term neonates and adults (mutational frequency 0.8% and 1.8%). We conclude that preterm infants have no limitations in gene segment usage, whereas the diversity of CDR3 is restricted throughout gestation.

scholarly journals Individual BFU-E in polycythemia vera produce both erythropoietin dependent and independent progeny

Blood ◽  
1983 ◽  
Vol 61 (5) ◽  
pp. 876-884 ◽  
Author(s):  
J Cashman ◽  
D Henkelman ◽  
K Humphries ◽  
C Eaves ◽  
A Eaves
Keyword(s):  
Polycythemia Vera ◽  
Peripheral Blood ◽  
Small Volume ◽  
Early Stage ◽  
Final Concentration ◽  
The Other ◽  
Normal Individuals ◽  
Additional Control ◽  
Minor Population ◽  
A Minor

Abstract Erythropoietic progenitors from peripheral blood of normal individuals or patients with polycythemia vera (PV) were cultured in methylcellulose medium containing 2.5 U/ml of erythropoietin (Ep). After 7–9 days, colonies considered to be early stage large bursts were individually removed, resuspended in a small volume of fresh methylcellulose medium, and then divided between 2 dishes. To one of these secondary cultures, sufficient Ep was added to bring the concentration of Ep up to approximately 3 U/ml. To the other was added an equal volume of medium but no Ep. The final concentration of Ep in these cultures was determined to be less than 0.01 U/ml. Nine days later, both types of secondary cultures were scored for the presence of colonies containing 8 or more hemoglobinized erythroblasts. Of 90 primary colonies from 3 normal individuals assessed in this way, 59 gave secondary erythroid colonies in the high Ep cultures, while none gave secondary erythroid colonies in the low Ep cultures. Additional control experiments in which primary colonies from normal individuals were divided into duplicate high Ep cultures showed that on average, the procedure used divided primary colonies equally. Of 109 primary colonies from 5 PV patients that yielded secondary erythroid colonies in the high Ep cultures, 21 yielded no secondary erythroid colonies in the low Ep cultures. The other 88 yielded erythroid colonies in both, but the secondary colonies in the low Ep cultures were consistently smaller in size and significantly fewer in number. Similar results were obtained when primary colonies were generated in cultures to which no Ep was added. These findings indicate that primitive BFU-E in patients with PV can be subdivided into 2 populations: a minor population restricted to the production of erythroid colony-forming cells (Ep- dependent progenitors) that require Ep for their detection, and a major population that is not restricted in this way. In addition, these experiments show that most of the primitive BFU-E that generate Ep- independent progenitors also produce significant numbers of cells that are Ep-dependent.

scholarly journals IgM+IgD+CD27+ B cells are markedly reduced in IRAK-4–, MyD88-, and TIRAP- but not UNC-93B–deficient patients

Blood ◽  
2012 ◽  
Vol 120 (25) ◽  
pp. 4992-5001 ◽  
Author(s):  
Sandra Weller ◽  
Mélanie Bonnet ◽  
Héloïse Delagreverie ◽  
Laura Israel ◽  
Maya Chrabieh ◽  
...  
Keyword(s):  
B Cells ◽  
B Cell ◽  
Signaling Pathways ◽  
Size Distribution ◽  
Heavy Chain ◽  
Peripheral Blood ◽  
Somatic Hypermutation ◽  
Key Factors ◽  
Tlr Signaling ◽  
B Cell Subsets

Abstract We studied the distribution of peripheral B-cell subsets in patients deficient for key factors of the TLR-signaling pathways (MyD88, TIRAP/MAL, IL-1 receptor–associated kinase 4 [IRAK-4], TLR3, UNC-93B, TRIF). All TLRs, except TLR3, which signals through the TRIF adaptor, require MyD88 and IRAK-4 to mediate their function. TLR4 and the TLR2 heterodimers (with TLR1, TLR6, and possibly TLR10) require in addition the adaptor TIRAP, whereas UNC-93B is needed for the proper localization of intracellular TLR3, TLR7, TLR8, and TLR9. We found that IgM+IgD+CD27+ but not switched B cells were strongly reduced in MyD88-, IRAK-4-, and TIRAP-deficient patients. This defect did not appear to be compensated with age. However, somatic hypermutation of Ig genes and heavy-chain CDR3 size distribution of IgM+IgD+CD27+ B cells were not affected in these patients. In contrast, the numbers of IgM+IgD+CD27+ B cells were normal in the absence of TLR3, TRIF, and UNC-93B, suggesting that UNC-93B–dependent TLRs, and notably TLR9, are dispensable for the presence of this subset in peripheral blood. Interestingly, TLR10 was found to be expressed at greater levels in IgM+IgD+CD27+ compared with switched B cells in healthy patients. Hence, we propose a role for TIRAP-dependent TLRs, possibly TLR10 in particular, in the development and/or maintenance of IgM+IgD+CD27+ B cells in humans.

scholarly journals Ig heavy chain CDR3 size diversities are similar after conventional peripheral blood and ex vivo expanded hematopoietic cell transplants

2001 ◽  
Vol 27 (4) ◽  
pp. 413-424 ◽  
Author(s):  
E Gokmen ◽  
C Bachier ◽  
FM Raaphorst ◽  
T Muller ◽  
D Armstrong ◽  
...  
Keyword(s):  
Heavy Chain ◽  
Peripheral Blood ◽  
Ex Vivo ◽  
Hematopoietic Cell ◽  
Cell Transplants ◽  
Ig Heavy Chain

scholarly journals Peripheral blood lymphocyte counts during standard conformal radiotherapy and TomoTherapy/IMRT for prostate cancer

2008 ◽  
Vol 7 (4) ◽  
pp. 223-227 ◽  
Author(s):  
M. Elsworthy ◽  
P.N. Plowman
Keyword(s):  
Peripheral Blood Lymphocyte ◽  
Peripheral Blood ◽  
Blood Lymphocyte ◽  
Small Volume ◽  
Intensity Modulated Radiation Therapy ◽  
Conformal Radiotherapy ◽  
Peripheral Lymphocyte ◽  
Prostate Radiotherapy ◽  
The Mean ◽  
Lymphocyte Populations

AbstractLymphopaenia is the earliest and the most sensitive routinely assessed biological parameter of corporeal radiation exposure in clinical practice; bone marrow, lymph nodes and peripheral blood lymphocyte populations are also at risk. During radical prostate radiotherapy, in 28 patients, the mean peripheral lymphocyte count fell from 1.76 × 109/l (standard deviation (SD) 0.63, 95% confidence interval (conf.) 0.23) to 1.10 × 109/l (SD 0.38, conf. 0.14), (p < 0.05). The question was asked as to whether intensity-modulated radiation therapy (IMRT) by TomoTherapy would cause more lymphopaenia than three-field conformal radiotherapy, bearing in mind the ‘low dose bath’ effect of IMRT and the long ‘beam-on’ times. Thirteen patients receiving three-field conformal radiotherapy experienced a fall in peripheral lymphocyte counts from 2.02 (SD: 0.62. conf. 0.43) to 1.17 × 109/l (SD: 0.47, conf. 0.26) after 34–38 Gy, as compared to a fall from 1.6 × 109/l (SD: 0.6, conf. 0.35) to 1.04 × 109/l (SD: 0.3, conf. 0.15) for 15 TomoTherapy patients—non-significant differences. We conclude that for this (approximately) standard, small-volume pelvic radiotherapy and to the dose under scrutiny, we cannot detect differences between the two radiotherapy techniques in terms of the lymphopaenia accruing. Neutrophil counts were similarly non-significantly different.

Successful Customization of Peripheral Blood Stem Cell Harvest Volume and Cell Concentration during Apheresis.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4992-4992 ◽  
Author(s):  
Michael J. Watts ◽  
Stuart J. Ings ◽  
Carmen Balsa ◽  
Caroline Penn ◽  
David Leverett ◽  
...  
Keyword(s):  
Stem Cell ◽  
Peripheral Blood ◽  
Healthy Donor ◽  
Small Volume ◽  
Peripheral Blood Stem Cell ◽  
Autologous Plasma ◽  
Cell Counts ◽  
Liquid Storage ◽  
Healthy Donors ◽  
Harvest Volume

Abstract The high white cell counts associated with peripheral blood stem cell (PBSC) harvests make these products particularly prone to cellular damage during storage in the liquid phase. We recently described unexpectedly high thaw clonogenic losses of PBSC stored overnight at 4 deg C prior to cryopreservation. This was associated with high harvest white cell counts and delayed engraftment in a cohort of patients receiving autologous transplantation procedures (Watts et al 2003 Blood 102:113 abstract 127). We showed in this study that pre-diluting the cells in autologous plasma to a WBC of 100x10^9/L preserved clonogenic yields during liquid storage and post freeze-thaw and suggested an upper WBC threshold of 200x10^9/L for liquid storage. In many patients however, to achieve this count or below would require further dilution of the cells. Conversely, where cells are to be frozen immediately, it is advantageous to collect a low PBSC harvest volume to fully utilize cryostorage space and to achieve this without centrifugation. The present study demonstrates that the collection of either a low white count or low volume PBSC harvest can be controlled successfully using the GAMBRO BCT Spectra AutoPBSC apheresis machine (version 6.1) and avoids the need for any further laboratory manipulations. This machine allows the adjustment of the amount of autologous plasma used to rinse each cycle of PBSC cells into the collection bag and is referred to as the “plasma chase volume”. The plasma chase volume was set to maximum (20ml/cycle) for healthy donor harvests for shipping, and to minimum (4ml/cycle) where the cells were for immediate cryopreservation. A total of 114 harvests from 99 mobilised healthy donors were collected using the maximum plasma chase volume whereas 527 autograft harvests from 365 mobilised patients were collected using the lowest plasma chase setting. The median (range) WBC and volume of the 114 healthy donor harvests was 100 (28–174)x10^9/L and 473 (54–871) ml respectively. The median (range) WBC and volume of the 527 “small volume” harvests for cryopreservation was 254 (51–495)x10^9/L and 66 (20–180) ml respectively. To determine whether the maximal plasma chase setting affected the progenitor dose collected, we compared the first day harvest of the 99 mobilised healthy donors obtained with the Spectra autoPBSC with that from 114 healthy donors collected on the standard manual Spectra (n=63) and CS3000 (n=51) apheresis machines. The median (range) CD34+ cell yield was 361 (34–1,380), 291 (21–1,356) and 259 (37–738)x10^6 respectively. The first day median (range) CD34+ cell yield x10^6 of the 365 mobilised patients where small volume autograft harvests were collected on the Spectra AutoPBSC was 202 (0–7,569) compared to 195 (0–5,054) using the manual Spectra (n=142) and 152 (0–4,830) x10^6 using the CS3000 machine (n=813). Our policy is to dilute any harvest for storage/shipping with a nucleated cell count greater than 200x10^9/L with autologous plasma, but none of the donor harvests exceeded this threshold and no laboratory manipulation was required. In the case of the autograft harvests for immediate cryopreservation, 502/527(95%) of the collections were 100ml or less. In conclusion, this study demonstrates for the first time that the cell count and volume of the PBSC harvest required can be customized at apheresis, that this is not detrimental to progenitor yields and results in a product that is optimal for storage/shipping without laboratory intervention.

The Immunoglobulin G Heavy Chain Repertoire in Multiple Sclerosis Plaques Is Distinct from the Heavy Chain Repertoire in Peripheral Blood Lymphocytes

2001 ◽  
Vol 98 (2) ◽  
pp. 258-263 ◽  
Author(s):  
Gregory P. Owens ◽  
Mark.P. Burgoon ◽  
Jacqueline Anthony ◽  
Bette K. Kleinschmidt-DeMasters ◽  
Donald H. Gilden
Keyword(s):  
Multiple Sclerosis ◽  
Heavy Chain ◽  
Immunoglobulin G ◽  
Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Blood Lymphocytes

scholarly journals Individual BFU-E in polycythemia vera produce both erythropoietin dependent and independent progeny

Blood ◽  
1983 ◽  
Vol 61 (5) ◽  
pp. 876-884 ◽  
Author(s):  
J Cashman ◽  
D Henkelman ◽  
K Humphries ◽  
C Eaves ◽  
A Eaves
Keyword(s):  
Polycythemia Vera ◽  
Peripheral Blood ◽  
Small Volume ◽  
Early Stage ◽  
Final Concentration ◽  
The Other ◽  
Normal Individuals ◽  
Additional Control ◽  
Minor Population ◽  
A Minor

Erythropoietic progenitors from peripheral blood of normal individuals or patients with polycythemia vera (PV) were cultured in methylcellulose medium containing 2.5 U/ml of erythropoietin (Ep). After 7–9 days, colonies considered to be early stage large bursts were individually removed, resuspended in a small volume of fresh methylcellulose medium, and then divided between 2 dishes. To one of these secondary cultures, sufficient Ep was added to bring the concentration of Ep up to approximately 3 U/ml. To the other was added an equal volume of medium but no Ep. The final concentration of Ep in these cultures was determined to be less than 0.01 U/ml. Nine days later, both types of secondary cultures were scored for the presence of colonies containing 8 or more hemoglobinized erythroblasts. Of 90 primary colonies from 3 normal individuals assessed in this way, 59 gave secondary erythroid colonies in the high Ep cultures, while none gave secondary erythroid colonies in the low Ep cultures. Additional control experiments in which primary colonies from normal individuals were divided into duplicate high Ep cultures showed that on average, the procedure used divided primary colonies equally. Of 109 primary colonies from 5 PV patients that yielded secondary erythroid colonies in the high Ep cultures, 21 yielded no secondary erythroid colonies in the low Ep cultures. The other 88 yielded erythroid colonies in both, but the secondary colonies in the low Ep cultures were consistently smaller in size and significantly fewer in number. Similar results were obtained when primary colonies were generated in cultures to which no Ep was added. These findings indicate that primitive BFU-E in patients with PV can be subdivided into 2 populations: a minor population restricted to the production of erythroid colony-forming cells (Ep- dependent progenitors) that require Ep for their detection, and a major population that is not restricted in this way. In addition, these experiments show that most of the primitive BFU-E that generate Ep- independent progenitors also produce significant numbers of cells that are Ep-dependent.

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