96. Jahrestagung der Deutschen Gesellschaft für Pathologie e. V ...

Abstracts
FR-P-146
Pneumocystis jirovecii: value of a standardized diagnostic procedure
with molecular pathology combined with cytochemistry
T . Zeiske 1 , A . Schad 1 , T . Wehler 2 , E . Springer 1 , A .J . Ullmann 2 , C .J . Kirkpatrick 1 ,
T . Hansen 1
1 University of Mainz, Institute of Pathology, Mainz, 2 University of Mainz,
Third Department of Internal Medicine, Mainz
Aims. Pneumocystis jirovecii (PJ) is a frequent cause of pulmonary infection
in patients with the acquired immunodeficiency syndrome and
other immunosuppressive conditions. Often PJ pneumonia represents
a life-threatening disorder requiring a sensitive and fast diagnosis. We
report on our experience on a combined diagnostic procedure for the
detection of PJ.
Methods. A total number of 602 cytological specimens were studied between
2008 and 2010 at the University Medical Center Mainz. As a standard
approach, all specimens have been routinely analyzed by Grocott’s
silver stain and nested-polymerase chain reaction (PCR) simultaneously.
The frequency of positive PJ results revealed by both methods was then
compared with respect to two different sampling procedures [bronchoalveolar
lavage (BAL) and sputum specimen]. For a subgroup of 18 patients,
we analyzed the clinical course over a one-year period.
Results. Our cohort included 441 BAL (73.3%) and 161 sputum specimens
(23.7%). In BAL, we found 22.45% of cases positive for PJ (n=99), with
24.2% of these diagnosed by both cytochemistry and PCR, whilst 75.8%
were detected by PCR alone. In sputum specimens, 33 PJ-positive cases
could be found (20.5%), most of them (28/33) being detected by PCR
alone. In about 24% of all specimens evaluated, cytochemistry revealed
various types of fungi such as candida and aspergillus. In the subpopulation
examined for the clinical course, we found 14/18 patients requiring
ventilation by respirator. In that PJ group, the large majority (i.e. 11/14)
was detected solely by PCR.
Conclusions. For the diagnosis of clinically relevant cases with PJ, the
combination of PCR with cytology/cytochemistry is mandatory. It remains
to be investigated, whether additional detection of fungi (by cytochemistry)
influences the clinical outcome of PJ patients.
Poster: Hämatopathologie
FR-P-147
Bone marrow biopsies of patients with haematopoietic and lymphoid
disorders – epidemiology, chromosomal aberrations and
molecular pathology
S . Hehne1 , S .M . Schulze1 , P . Richter1 , C . Geier1 , Y . Chen1 , A . von Deimling 2 ,
I . Petersen1 1 2 Jena University Hospital, Heidelberg University Hospital, Institute of
Neuropathology
Aims. Bone marrow biopsy of the iliac crest is the first and most important
step in the diagnostics of haematopoietic disorders.
Methods. The biopsies of the years 2006 and 2007 from the institute of
pathology of the Jena university hospital were retrospectively analyzed
for clinicopathological parameters. In addition, the Mitelman database
was retrieved for chromosomal aberrations.
Results. The analysis of 2820 reports from 1185 patients revealed that lymphomas,
plasmocytoma and acute leukaemia were most frequent. Males
predominated in myeloproliferative neoplasms and lymphoma subtypes,
particularly CLL, except for plasmocytoma and acute leukaemia. A
peak incidence was seen between 61 and 70 years of age with a varying
pattern for single entities. The database search revealed that ALL, AML,
CLL and cmL were mainly diploid while Hodgkin lymphoma, mature
B-cell lymphoma and multiple myeloma mostly carried hyperdiploid
chromosome numbers. Numerical aberrations like chromosome 8 gains
130 | Der Pathologe · Supplement 1 · 2012
in hyperdiploid cmL were prominent in specific subgroups. Molecular
testing is exemplified in cmL, plasma cell myeloma and hairy cell leukaemia.
Conclusions. The study highlights typical clinicopathological characteristics
and new genetic findings in haematopoietic and lymphoid neoplasms
with relevance for the new WHO classification and beyond. We
hope that it may help in the differential diagnosis of bone marrow biopsies.
FR-P-148
Clonally related nodular lymphocyte-predominant Hodgkin lymphoma
and classical Hodgkin lymphoma occurring as a collision
lymphoma
M . Szczepanowski1 , N . Masqué-Soler1 , I . Oschlies1 , W . Schmidt 2 , A . Lück3 ,
W . Klapper1 1University Hospital Campus Kiel/Institute of Pathology, Section Hematopathology,
House 14 , Kiel, 2Pathology Practice, Rostock, 3Practice for Internal
Medicine, Hematology and Oncology, Rostock
Aims. Nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL)
with the typical lymphocyte-predominant (LP) cells, and classical Hodgkin
lymphoma (cHL), characterized by Hodgkin and Reed-Sternberg
(HRS) cells, are considered two distinct diseases whose co-occurrence in
one patient is extremely rare. We report on clonal relatedness in a case of
concurrent NLPHL and cHL, residing in one lymph node in a 48-yearold
male patient.
Methods. We diagnosed synchronous NLPHL (CD20+, CD30−, LMP1−)
and cHL (CD30+, CD20−, LMP1−) in formalin-fixed paraffin-embedded
(FFPE) tissue sections of one lymph node in a 48-year-old male patient.
CD20 or CD30 labeled neoplastic cells were separately collected by laserassisted
microdissection (LCM). Immunoglobulin heavy chain (IGH)
gene rearrangements were pre-amplified by standard multiplex polymerase
chain reaction (PCR) and subjected to fragment analyses and
Sanger sequencing.
Results. In the frame work (FR) 3 multiplex PCR, fragments of 120/121
and 128 base pairs (bp) in LP cells and 120/121 bp in HRS cells were obtained.
The 120/121 bp fragment shared by both LP and HRS cells was
re-amplified by VH1 and JHrev primers as a clear 121 bp fragment.
Sequencing analyses of the shared 121 bp product revealed an identical
rearrangement consisting of IGHV1-2*3/IGHD4-17*01/IGHJ4*02
with identical VH-DH junction (5’-GCCAT-3’) and DH-JH junction
(5’-CCTCTCTTTTGACTG-3’) sequences and a mutation rate of 5.6%
in both entities. Consequently, PCR with allele-specific oligonucleotides
(ASO) yielded identically sized fragments in both LP and HRS cells.
Conclusions. We conclude that both approaches, sequencing of VH1/
JHrev PCR products and amplification of identically sized ASO products,
are highly indicative for a clonal relationship of the concurrent
NLPHL and cHL. The findings point to a common precursor B-cell of
germinal center origin for both, the cHL and the NLPHL, in analogy
to well documented clonal relations in cases of concurrent or sequential
Hodgkin lymphoma (HL) and non-HL.