Haptoglobin is a long known acute phase protein involved in hemoglobin breakdown, which was thought to be from the liver. The discovery of pHp was achieved by applying transcriptome analysis to human lung tissues which were subjected to a short term culture and infected or not infected with Haemophilus influenzae. These tissues were fixed and paraffin-embedded by the HOPE-technique.

Background: The acute-phase protein haptoglobin (Hp) and its receptor CD163 serve as immunomodulators and possess anti-inflammatory besides antioxidant functions. Objectives: To further understand the role of the recently described pulmonary Hp (pHp) and its receptor CD163 in case of inflammation and infection, pHp and CD163 were investigated on mRNA and protein level to gain insight into the cellular events taking place upon stimulation with the inflammatory mediators LPS, Pam3, cytokine IL-6 and dexamethasone, and upon infection with respiratory pathogens (Haemophilus influenzae, Streptococcuspneumoniae and Chlamydia pneumoniae) by use of a human ex vivo tissue culture model and cell cultures of A549 and alveolar epithelial cells type II. In addition, pHp and CD163 expression in COPD and sarcoidosis was assessed. Methods: We conducted experiments using 942 ex vivo cultured lung samples applying immunohistochemistry, immunocytochemistry, in situ hybridization, immunofluorescence, real-time PCR, RT-PCR, slot and Western immunoblot analyses with tissue lysates and culture supernatants as well as ELISA and cytometric bead array analyses. Results: This study describes for the first time the expression, regulation and secretion of pHp and its receptor CD163 in the human lung. The release of soluble mediators from A549 cell line and human monocyte-derived macrophages was observed indicating that Hp differentially activates the release of soluble mediators and major chemoattractants. Conclusions: The findings indicate a native function of pHp and CD163 as functional pulmonary defense elements due to local expression, regulation and secretion during lung infection and as part of the inflammatory immune response of the respiratory system.

In the last decade, pathologic approaches concerning diagnosis and treatment of lung carcinomas have increasingly moved towards the implementation of molecular methods into the process of decision. In this study, an overview is given referring to the variety of tumors in the lung including common primary lung neoplasms and secondary tumors, and a modus operandi is presented which integrates immunology as well as molecular pathology within the process of finding correct diagnoses. Besides the conventional and approved methods and techniques leading to appropriate treatment including so-called targeted therapies, pathologist's work meanwhile depends on both histologic and molecular results. Since molecular techniques have increasingly entered the field of routine diagnostics, challenges and possibilities have changed and are still rapidly developing. The proceeding integration of molecular-biologic investigations into the process of diagnosing has changed the nature of diagnostics and will continuously grow in the near future. Only by obtaining a proper diagnosis, the optimal treatment of a patient can be assured, whereupon the knowledge of gene mutations and/or altered protein expression is crucial. By identifying those novel molecular target structures, the therapeutic spectrum is tremendously enlarged and will finally improve the patient's prognosis by personalized targeted therapies.

Einleitung: Untersuchungen am Proteom humaner Lungengewebe setzen eine adäquate Bearbeitung der Gewebe voraus. Essentiell für das Ergebnis aller Proteinuntersuchungen an Archivmaterial ist die Qualität der Gewebefixierung mit wenig molekularen Alterationen auch nach längerer Lagerungszeit sowie die Möglichkeit, die fixierten Gewebe über Jahre archivieren zu können. Die dafür zur Verfügung stehenden Standard-Techniken sind das Einfrieren der Gewebe in flüssigem Stickstoff (und Lagerung bei -80°C) oder die Formalinfixierung mit folgender Paraffineinbettung. Beide Methoden gehen mit der Entstehung von Artefakten einher: Formalin ist für Denaturierung und Quervernetzung von Proteinen verantwortlich und wird vornehmlich bei histologischen Untersuchungen verwendet, Gefriermaterial als Standardfixierung für Proteomstudien besitzt eine schlechte Morphologie und ist eingeschränkt in der Dauer seiner Lagerfähigkeit (ab 3 Jahre). Methoden: In dieser Studie wurden Proteomuntersuchungen mithilfe von 2-D-Elektrophorese und Massenspektrometrie auf Basis der HOPE (Hepes-glutamic acid buffer mediated organic solvent protection effect)-Technik etabliert. Dies bietet Vorteile in Fixierungs- und Archivierungsqualität und gewährleistet guten morphologischen wie molekularen Erhalt, da keine Quervernetzung der Proteine und kaum Denaturierung stattfindet. Die HOPE-Technik wird mit einer Paraffineinbettung abgeschlossen. Zur Anwendung kamen humane Lungenkarzinomgewebe und tumorfreie Lungengewebe. Ergebnisse: Mit HOPE-Material konnten reproduzierbare Spotmuster sowie eine erfolgreiche Proteinidentifikation über Massenspektrometrie erreicht werden. Die hohe Auflösung der Muster und die Anzahl detektierbarer Proteine sind mit denen von Gefriermaterial vergleichbar. Diskussion: Diese Methode bietet neue Möglichkeiten bei Studien an Paraffin-Gewebearchiven mit dem Ziel der Identifikation krankheitsspezifischer Proteinexpressionsmuster und einzelner krankheitsrelevanter Moleküle.

Introduction: The acute phase protein Haptoglobin (Hp) consists of two different polypeptide chains. Originating from the liver; the biological function is binding free hemoglobin (Hb) and preventing oxidative stress. Elevated amounts of Hp in plasma were observed were infection, inflammation and malignant diseases. Purpose of this study was to investigate protein and mRNA expression within human lung tissues and tumors. Methods: We performed immunohistochemistry, in situ hybridization and RT-PCR analyzing 115 tissue samples. 47 adenocarcinomas, 42 squamous cell carcinomas, 13 small cell lung cancers and 13 tumor-free lungs were investigated. After lobectomy/pneumonectomy tissues were fixed and paraffin-embedded using formalin (immunohistochemistry) or the HOPE-solution (other methods). Results: Immunohistochemistry and in situ hybridization revealed a high level of Hp expression in adenocarcinomas (40.4%) in contrast to squamous cell carcinomas (4.8%, expression in alveolar epithelial cells type II surrounding the tumor). One small cell carcinoma showed Hp expression. In tumor-free lungs we located Hp in alveolar macrophages; alveolar epithelial cells type II and bronchi. RT-PCR results confirmed elevated expression. Discussion: We characterized that Hp protein expressed in lung cancers and tumor-free lungs showing an extra hepatic function. Due to the known immunosuppressive properties of Hp, its broad synthesis is strongly suggestive for a function as a fundamental, pulmonary, local defense element and confirms recent reports that haptoglobin plays an important role in protection processes and repairing mechanisms of injured tissues.