Towards a commercial process for the manufacture of genetically modified T cells for therapy.

The recent successes of adoptive T-cell immunotherapy for the treatment of hematologic malignancies have highlighted the need for manufacturing processes that are robust and scalable for product commercialization. Here we review some of the more outstanding issues surrounding commercial scale manufacturing of personalized-adoptive T-cell medicinal products. These include closed system operations, improving process robustness and simplifying work flows, reducing labor intensity by implementing process automation, scalability and cost, as well as appropriate testing and tracking of products, all while maintaining strict adherence to Current Good Manufacturing Practices and regulatory guidelines.

Expression of gangliosides GM3 (NeuAc) and GM3 (NeuGc) in myelomas and hybridomas of mouse, rat, and human origin.

In this study gangliosides from various myelomas and hybridomas of mouse, rat, and human origin were characterized by thin-layer and high-performance liquid chromatography, immunological methods (overlay technique) and fast atom bombardment mass spectrometry. Exclusively GM3 substituted with C24:1- and C16:0-fatty acid, was found in all B cell-derived cell lines. C18 sphingosine was the single long chain base in each GM3 ceramide portion.

Rejection prophylaxis with interleukin-2 receptor antibody BT 563: mechanisms of action on human cells.

We investigated the in vitro immunosuppressive effect of BT 563, a monoclonal antibody, against the alpha-chain of the human interleukin-2 (IL-2) receptor (p 55), which has been used to prevent transplant rejection in several clinical trials. We also measured the proliferative T cell alloresponse and pCTL frequencies of BT563-treated kidney transplant patients. In mixed lymphocyte cultures BT 563 caused a reduction of T cell proliferation to about 50%.

[Lung imaging in intensive medicine. A comparison of digital storage-screen imaging and conventional film-screen imaging].

Luminescence radiography is a new digital imaging modality, which can be used instead of conventional film radiography for many examinations. Imaging capabilities of luminescence radiography were compared with film radiography by test images and examinations especially in intensive care radiology. The results demonstrate, that luminescence radiography is an advantageous imaging method for intensive care radiology.

[Cerebral blood flow scintigraphy using 99mTc-HM-PAO in the diagnosis of brain death].

In 9 patients with the clinical diagnosis of brain death, a HM-PAO-SPECT was performed to document the arrest of cerebral circulation. Besides brain-stem areflexia, negative apnea-test and an observation time of at least 12 hours neurophysiological examinations (EEG and evoked potentials) were performed. In 4 cases, an additional arterial digital subtraction angiography proved the arrest of cerebral circulation shown in SPECT.

Forssman glycolipid, an antigenic marker for a major subpopulation of macrophages from murine spleen and peripheral lymph nodes.

Three hybridoma clones were isolated after hybridization of a mouse myeloma line with splenocytes from rats immunized with Forssman glycosphingolipid (Fo). Two of these clones produced Fo-specific monoclonal antibodies (MAB) of the IgM class, one MAB of the IgG2c class. In complement-dependent depletion experiments and immunofluorescence studies on the nature of Fo-positive leukocytes in CBA/J mice the following results were obtained: whereas blood monocytes, polymorphonuclear leukocytes, and lymphocytes were Fo negative, 5 to 10% of suspended spleen cells were positive.

An improved semi-quantitative enzyme immunostaining procedure for glycosphingolipid antigens on high performance thin layer chromatograms.

An immunoassay is described which allows the detection of glycosphingolipid (GSL) antigens on high performance thin layer chromatograms (HPTLC). The method involves: (1) the separation of GSL on HPTLCs; (2) incubation with specific antibodies against carbohydrate structures of GSL, and (3) the detection of specifically bound antibodies with alkaline phosphatase-conjugated second antibodies and 5-bromo-4-chloro-3-indolyl-phosphate (BCIP) as substrate. Using a monoclonal rat IgG2c antibody against Forssman GSL, a BALB/c monoclonal antibody against asialo GM2, and polyclonal rabbit antibodies against asialo GM1, it was shown that as little as 3 ng GSL antigen could be detected in a procedure taking detected in a procedure taking only 4 h to perform.

The glycosphingolipid globoside as a serological marker on cytolytic T lymphocyte precursors and alloantigen-responsive proliferating T lymphocytes in murine spleen.

Biochemical analyses of murine lymphocytes have shown that the glycosphingolipid globoside (Glo) is present exclusively on alloantigen-stimulated murine T lymphocytes (Gruner, K. R., Van Eijk, R.

[Anesthesia in concomitant bronchopulmonary diseases].

The goal of any anaesthesiologist delivering anaesthesia to patients with pre-existing lung disease has to be the prevention of intra- and postoperative complications. Changes in gas exchange imposed by anaesthesia and surgery can cause dangerous disturbances in oxygen transport and in acid-base balance. Uptake and elimination of volatile and gaseous anaesthetics depend upon normal respiratory function.

[The need for a specific postoperative, total parenteral nutrition therapy after various intra-abdominal procedures].

119 metabolically healthy surgical patients, who had to undergo elective intraabdominal surgery, were separated into upper -- cholecystectomy, gastric resection -- and lower -- colonic and sigma surgery -- intraabdominal procedures and divided into 8 groups with different infusion regimes. During the early postoperative period patients with upper abdominal surgery had significantly higher postoperative enzyme changes compared with patients with lower abdominal surgery. Patients with cholecystectomy had the significantly highest postoperative enzyme changes.

[Comparison of xylose and glucose as energy sources in hypocaloric, postoperative parenteral nutrition].

After a 5 day preoperative preparing period 24 metabolically healthy patients, who had to undergo gastric resection, were fed postoperatively by hypocaloric total parenteral nutrition for a 5 day period. Group I (n = 13) received 0,11 g Xylitol/kg BW X h; Group II (n = 11) received 0,11 g Glucose/kg BW X h; Both groups received 1,71 g L-crystalline amino acids/kg BW X day. During the whole postoperative period group II had significantly higher serum Glucose and Insulin levels.

[Psychosomatic effects of intramuscular premedication wtih flunitrazepam, innovar and a placebo in combination with atropine (author's transl)].

A double-blind technique was used for studying the psychic and somatic effects of premedication with flunitrazepam, innovar and a placebo in 116 patients. Innovar and the placebo produced the same results. Flunitrazepam reduced anxiety and apprehension and had a significantly higher amnesic effect.

[Differences in postoperative metabolism after pre- and postoperative beginning of total parenteral nutrition].

In this study we intended to investigate the influence of a 24-h-preoperative total parenteral nutrition (TPN) therapy--10 surgical patients, group 1--compared with the postoperative beginning of TPN--9 surgical patients, group 2--on the postoperative metabolism. Most probably due to preoperatively depleted glycogen storages in G 2, the urinary carbohydrate losses declined more rapidly postoperatively compared with G 1. 45 mMol/l of sodium did not cover the postoperative requirements of G 1.

[Postoperative metabolism--differences between pre- and postoperative start of total parenteral nutrition (author's transl)].

We investigated the postoperative metabolism of patients undergoing gastric operation by beginning total parenteral nutrition pre- (group 1) and postoperatively (group 2). A third group remained fasting until 3 h after the surgical intervention. Because of the high serum glucose concentrations and the high glucose losses in the 24-h-urine on the operation day in group 1 and 2 only the basic glucose requirement (150--250 mg/24h) should be given in the early postoperative period.

[Differences in postoperative metabolism in reference to pre- and postoperative initiation of total parenteral nutrition. I].

We investigated the postoperative metabolism of patients undergoing gastric resection by beginning total parenteral nutrition pre- (group 1) and postoperatively (group 2). A third group remained fasting until 3 h after the surgical intervention. Because of the high serum glucose concentrations and the high glucose losses in the 24-h urine on the operation day in group 1 and 2 only the base glucose requirement (150--250 g/24/h) should be given in the early postoperative period.

[The sedative and amnesic effects of flunitrazepam during regional anaesthesia (author's transl)].

The sedative, hypnotic and amnesic effects of flunitrazepam, a new benzodiazepine, were tested in doses of 0 (placebo), 0,4 mg and 0,8 mg. These doses were sufficient to produce a satisfying sedative effect on patients, being operated on in regional anaesthesia. In few cases we found undesirable effects.